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Latest revision as of 11:14, 27 October 2013

Notebook: June

03.06.2013

Digest

Investigator: Franzi

Aim: Construction of P_NR-test-vector → Digestion of combined 2 BioBricks.

10 µl DNA template
0,5 µl enzyme 1
0,5 µl enzyme 2
2 µl Cut Smart buffer
7 µl bidest. H2O

Samples:

Digestion of pSB1A3 iBB1+5 with XbaI and PstI-HF

Digestion of pSB1A3 iBB6+3 with EcoRI-HF and SpeI-HF.

The samples were incubated for 1 h at 37 °C.

They were purified via QIAgen Gel Extraction Kit (Qiagen). Elution into 20 µl bidest. H2O.

Ligation

Investigator: Franzi

Aim: Construction of P_NR-test-vector → Ligation of two combined 2 BioBricks in pSB1C3.

45 ng vector DNA (pSB1C3; EcoRI/PstI-cut)
140 ng iBB6+3 (EcoRI/PstI-cut)
150 ng iBB1+5 (EcoRI/PstI-cut)
2 µl 10x T4 DNA ligase buffer
1 µl T4 DNA ligase
ad 20 µl bidest. H2O

The samples were incubated for 1 h at RT.

Transformation

Investigator: Franzi

Aim: Construction of P_NR-test-vector → Transformation of combined 4 BioBricks in pSB1C3 into E. coli DH5α.

Complete ligation samples were transformed into E. coli DH5α (30 min on ice, 60 sec 42 °C, 60 min 37 °C + 900 µl LB), plated on LB_CM, overnight at 37 °C.

Ligation

Investigator: Franzi

Aim: Construction of BioBricks → Ligation of single BioBricks into pSB1C3.

45 ng vector DNA (pSB1C3; EcoRI/PstI-cut)
X ng insert
1 µl 10x T4 DNA ligase buffer
1 µl T4 DNA ligase
ad 10 µl bidest. H2O

Samples (all EcoRI/PstI-cut):

120 ng eGFP (iBB9)

60 ng SPTP1 (iBB10)

50 ng SPTP2 (iBB11)

55 ng SP1 (iBB12)

50 ng SP2 (iBB13)

The samples were incubated for 3 h at RT.

Transformation

Investigator: Franzi

Aim: Construction of BioBricks → Transformation of single BioBricks in pSB1C3 into E. coli DH5α.

Complete ligation samples were transformed into E. coli DH5α (30 min on ice, 60 sec 42 °C, 60 min 37 °C + 900 µl LB), plated on LB_CM, overnight at 37 °C.

Inoculation

Investigator: Franzi

Aim: Construction of P_fcpB-test-vector and antibody-vector → Inoculation of transformants for miniprep.

3 colonies of pSB1A3 iBB1+6 inoculated in 5 ml LB_amp, 4 colonies of pSB1C3 iBB4864 inoculated in 5 ml LB_CM, overnight at 37 °C.

04.06.2013

Miniprep

Investigator: Franzi

Aim: Construction of P_fcpB-test-vector and antibody-vector → Miniprep of pSB1A3 iBB1+6 and pSB1C3 iBB4864.

Miniprep of the liquid cultures (“QIAprep Spin Miniprep Kit” (Qiagen, Düsseldorf)), eluted in 30 µl bidest. H2O. 2 Cultures of pSB1C3 iBB4864 discarded (red colour).

Digest

Investigator: Franzi

Aim: Construction of P_fcpB-test-vector and antibody-vector → Control digestion of pSB1A3 iBB1+6 and pSB1C3 iBB4864.

1 µl DNA template
0,5 µl EcoRI-HF
0,5 µl PstI-HF
1 µl Cut Smart buffer
7 µl bidest. H2O

The samples were incubated for 2 h at 37 °C.

Gel electrophoresis

Gel substances

1% Agarose gel
10 µl RedSafe per 50 ml gel
6 µl 2-Log DNA Ladder (NEB)
6x loading buffer (for samples)

Lane	Content	Expectations
2+3	iBB4864 K1-2	2580 bp
4	empty
5-7	iBB16 K3-5	650 bp

Digest

Investigator: Franzi

Aim: Construction of P_fcpB-test-vector and antibody-vector → Digestion of combined 2 BioBricks in pSB1A3 and combined 4 BioBricks in pSB1C3.

10 µl DNA template
0,5 µl enzyme 1
0,5 µl enzyme 2
2 µl Cut Smart buffer
7 µl bidest. H2O

Samples:

Digestion of pSB1C3 iBB7631 K4 and pSB1A3 iBB16 K4 with XbaI and PstI-HF

Digestion of pSB1C3 iBB4864 K1 and pSB1A3 iBB54 K2 with EcoRI-HF and SpeI-HF.

The samples were incubated for 1 h at 37 °C.

They were purified via QIAgen Gel Extraction Kit (Qiagen). Elution into 20 µl bidest. H2O.

Ligation

Investigator: Franzi

Aim: Construction of P_fcpB-test-vector and antibody-vector → Ligation of two combined 2 BioBricks into pSB1C3 and two combined 4 BioBricks into pSB1A3.

Antibody:

80 ng vector DNA (pSB1A3; EcoRI/PstI-cut)
160 ng iBB7631
190 ng iBB4864
1 µl 10x T4 DNA ligase buffer
1 µl T4 DNA ligase
ad 10 µl bidest. H2O

Test-vector:

45 ng vector DNA (pSB1C3; EcoRI/PstI-cut)
150 ng iBB54
120 ng iBB16
1 µl 10x T4 DNA ligase buffer
1 µl T4 DNA ligase
ad 10 µl bidest. H2O

Both samples were incubated overnight at 16 °C.

Inoculation

Investigator: Franzi

Aim: Construction of P_NR-test-vector and BioBricks → Inoculation of transformants.

Transformations (03.06.13) inoculated in 5 ml LB_CM, overnight at 37 °C.

05.06.2013

Miniprep

Investigator: Franzi

Aim: Construction of P_NR-test-vector and BioBricks → Miniprep of single BioBricks and pSB1C3 iBB6315.

Miniprep of the liquid cultures (“QIAprep Spin Miniprep Kit” (Qiagen, Düsseldorf)), eluted in 30 µl bidest. H2O.

Digest

Investigator: Franzi

Aim: Construction of P_NR-test-vector and BioBricks → Control digestion of single BioBricks and pSB1C3 iBB6315.

1 µl DNA template
0,3 µl EcoRI-HF
0,3 µl PstI-HF
1 µl Cut Smart buffer
7,4 µl bidest. H2O

The samples were incubated for 1,5 h at 37 °C.

Gel electrophoresis

Gel substances

1% Agarose gel
10 µl RedSafe per 50 ml gel
6 µl 2-Log DNA Ladder (NEB)
6x loading buffer (for samples)

Lane	Content	Expectations
2-4	iBB9 K1-3	780 bp
5	empty
6-8	iBB10 K1-3	228 bp
		pSB1C3: 2 kb

Lane	Content	Expectations
2-4	iBB11 K1-3	147 bp
5	empty
6-8	iBB12 K1-3	123 bp
		pSB1C3: 2 kb

Lane	Content	Expectations
2-4	iBB13 K1-3	108 bp
5	empty
6-8	iBB6315 K1-3	1200 bp
		pSB1C3: 2 kb

Transformation

Investigator: Franzi

Aim: Construction of P_fcpB-test-vector and antibody-vector → Transformation of combined 4 BioBricks in pSB1C3 and combined 8 BioBricks in pSB1A3 into E. coli DH5α.

Ligations (04.06.13) were transformed into E. coli DH5α (30 min on ice, 60 sec 42 °C, 60 min (1C3)/40 min (1A3) 37 °C + 900 µl LB), plated on LB_CM (1C3)/LB_amp (1A3), overnight at 37 °C.

Sequencing

Investigator: Franzi

Aim: Construction of BioBricks → Examination of sequencing results of various constructs.

iBB9 K1 fw - AGB000K 263

iBB9 K1 rv - AGB000K 264

iBB9 K2 fw - AGB000K 265

iBB9 K2 rv - AGB000K 266

iBB10 K1 fw - AGB000K 267

iBB10 K2 fw - AGB000K 268

iBB11 K1 fw - AGB000K 269

iBB11 K2 fw - AGB000K 270

iBB12 K1 fw - AGB000K 271

iBB12 K2 fw - AGB000K 272

iBB13 K1 fw - AGB000K 273

iBB13 K2 fw - AGB000K 274

all correct, except iBB10 G→T pos.30, iBB11 T→C pos.24. Probably template mutated.

06.06.2013

Inoculation

Investigator: Franzi

Aim: Construction of P_fcpB-test-vector and antibody-vector → Inoculation of pSB1C3 iBB5416 and pSB1A3 iBB48647631 for miniprep.

3 colonies of pSB1C3 iBB5416 inoculated in 5 ml LB_CM, 4 colonies of pSB1A3 iBB48647631 inoculated in 5 ml LB_amp, overnight at 37 °C.

07.06.2013

Miniprep

Investigator: Patrick

Aim: Construction of P_fcpB-test-vector and antibody-vector → Miniprep of pSB1C3 iBB5416 and pSB1A3 iBB48647631.

Miniprep of the liquid cultures (“QIAprep Spin Miniprep Kit” (Qiagen, Düsseldorf)), eluted in 30 µl bidest. H2O.

Digest

Investigator: Patrick

Aim: Construction of P_fcpB-test-vector and antibody-vector → Control digestion of pSB1C3 iBB5416 and pSB1A3 iBB48647631.

1 µl DNA template
0,3 µl EcoRI-HF
0,3 µl PstI-HF
1 µl Cut Smart buffer
7,4 µl bidest. H2O

The samples were incubated for 1 h at 37 °C.

Gel electrophoresis

Gel substances

1% Agarose gel
10 µl RedSafe per 50 ml gel
6 µl 2-Log DNA Ladder (NEB)
6x loading buffer (for samples)

Lane

Content

Expectations

2-4

iBB5416 K1-3

pSB1C3: 2070 bp

iBB5416: 1500 bp

5-8

iBB48647631 K1-4

pSB1A3: 2150 bp

iBB48647631: 4157 bp

10.06.2013

Digest

Investigator: Lukas

Aim: Construction of P_NR-test-vector, P_fcpB-test-vector and antibody-vector → Digestion of pSB1A3 iBB48647631 and single BioBricks.

X µl DNA template
0,5 µl enzyme 1
0,5 µl enzyme 2
2,2 µl Cut Smart buffer
20 µl bidest. H2O

Samples:

Digestion of 1,5 µl pSB1A3 iBB48647631 with SpeI-HF and PstI-HF

Digestion of 5 µl pSB1C3 iBB5 and iBB9 with XbaI and PstI-HF

Digestion of 4 µl pSB1C3 iBB3 and iBB4 with EcoRI-HF and SpeI-HF.

The samples were incubated for 1 h at 37 °C.

They were purified via QIAgen Gel Extraction Kit (Qiagen). Elution into 20 µl bidest. H2O.

Ligation

Investigator: Christian

Aim: Construction of P_NR-test-vector, P_fcpB-test-vector and antibody-vector → Ligation of pSB1A3 iBB48647631 and single BioBricks in pSB1A3.

Ligation of pSB1A3 iBB48647631 with iBB5, pSB1A3 iBB48647631 with iBB3 and iBB9, pSB1A3 iBB48647631 with iBB4 and iBB9; overnight at 16°C

11.06.2013

Transformation

Investigator: Franzi

Aim: Construction of P_NR-test-vector, P_fcpB-test-vector and antibody-vector %rarr; Transformation of ligations.

Transformation of the ligations into E. coli DH5α (30 min on ice, 60 sec 42 °C, 25 min 37 °C + 900 µl LB), plated on LB_amp, overnight at 37 °C.

12.06.2013

Inoculation

Investigator: Franzi

Aim: Construction of P_NR-test-vector, P_fcpB-test-vector and antibody-vector → Inoculation of transformants for miniprep.

4 colonies of each transformation inoculated in 4 ml LB_amp, overnight at 37 °C.

13.06.2013

Miniprep

Investigator: Franzi

Aim: Construction of P_NR-test-vector, P_fcpB-test-vector and antibody-vector → Miniprep of transformants.

Miniprep of the liquid cultures (“QIAprep Spin Miniprep Kit” (Qiagen, Düsseldorf)), eluted in 30 µl bidest. H2O.

Digest

Investigator: Franzi

Aim: Construction of P_NR-test-vector, P_fcpB-test-vector and antibody-vector → Control digestion the minipreps and pSB1A3 iBB48647631 K3 (comparison of size).

1 µl DNA template
0,3 µl EcoRI-HF
0,3 µl PstI-HF
1 µl Cut Smart buffer
7,4 µl bidest. H2O

The samples were incubated for 1 h at 37 °C.

Gel electrophoresis

Gel substances

1% Agarose gel
10 µl RedSafe per 50 ml gel
6 µl 2-Log DNA Ladder (NEB)
6x loading buffer (for samples)

Lane	Content	Expectations
upper:
2+3	iBB49 K3-4	1260 bp
4-7	iBB39 K1-4	1030 bp
lower:
2	iBB49647631	4157 bp
3-6	iBB496476315 K1-4	4404 bp
7+8	iBB49 K1-2	1260 bp
		Vector pSB1A3: 2150 bp

IBB48647631 and iBB486476315 probably positive, but better repeat or do a Colony-PCR.

16.06.2013

Digest

Investigator: Lukas

Aim: Construction of antibody-vector → Repeat of control digestion.

1 µl DNA template
0,3 µl EcoRI-HF
0,3 µl PstI-HF
1 µl Cut Smart buffer
7,4 µl bidest. H2O

Templates: PSB1A3 iBB486476315 and pSB1A3 iBB48647631 K3 (comparison of size).

The samples were incubated for 1 h at 37 °C.

Gel electrophoresis

Gel substances

1% Agarose gel
10 µl RedSafe per 50 ml gel
6 µl GeneRuler™ 1kb plus DNA Ladder (Thermo Scientific)
6x loading buffer (for samples)

Lane	Content	Expectations
2+3	iBB48647631	4157 bp
4-7	iBB486476315	TBA bp
		Vector pSB1A3: 2150 bp

Too much DNA used, will be repeated as PCR.

17.06.2013

Digest

Investigator: Alex

Aim: Digest of pSB1A3_3+9 and pSB1A3_4+9 with SpeI and PstI, and corresponding inserts pSB1C3_5+4+1+6 and pSB1C3_6+3+1+5 with EcoRI and PstI.

1 µl DNA
0.3 µl Enzyme 1 (SpeI or EcoRI, resp.)
0.3 µl Enzyme 2 (PstI)
1 µl Cut Smart buffer
7.4 µl ddbidest. H2O

The samples were incubated for 1.5 h at 37 °C.

Approaches were unsewed by means of gel electrophoresis, corresponding bands were picked under UV, DNA extraction from gel was performed using DNA Gel Extraction Kit (QIAGEN).

Ligation, Transformation, Plasmid prep

Investigator: Alex

Aim: Assemble the combined bricks 5416 and 6315 into their appropriate vectors pSB1A3_39 and pSB1A3_49, resp.

Insert und plasmid were transformed using a 1:2 ratio. Samples were incubated ON at RT.
Ligation product were transformed into chemical competent DH5α cells (30 min on ice, 90 sec 42 °C, 45 min 37 °C + 900 µl LB).
Transformed cells were spread out on LB Amp culture plates.
Incubation ON at RT.
4 clones were picked for overnight at cultures.
A plasmid preparation was carried out the next day.

22.06.2013

Test digest

Investigator: Alex

Aim: Digest of pSB1A3_395416 and pSB1A3_496315 with EcoRI and PstI.

1 µl Plasmid DNA
0.3 µl EcoRI
0.3 µL PstI
1 µl Cut Smart
7.4 µl ddbidest. H2O

The samples were incubated for 2 h at 37 °C.

Gel electrophoresis

Gel substances

1% Agarose gel
10 µl Ethidium bromide 50 ml gel
4 µl 2-log DNA ladder (New England Biolabs)

Lane	Content	Expectations
upper:
2	iBB6315	1200 + 2000 bp
3	iBB49	1200 + 2000 bp
4-7	iBB496315 K4-1	2400 + 2000 bp
lower:
2	iBB5416	1500 + 2000 bp
3	iBB39	1000 + 2000 bp
4-7	iBB395416 K1-4	2400 + 2000 bp

We didn't receive all expected fragments. All expected fragments in lanes 4-7 were missing. Ligation failed ☹

24.06.2013

PCR

Investigator: Alex

Aim: Check right assembly of BioBrick BBa_K1071005 in the plasmid for antibody production

Volume	Reagent	Temp (°C)	Time
1 µl	pSB1A3_486476315	95	3 min
0.5 µl	Primer fwd	95	30 sec
0.5 µl	Primer rev	60	30 sec	x30
0.5 µl	dNTPs	72	42 sec
0.5 µl	Phusion polymerase	72	5 min
5 µl	Phusion buffer 5x	4	Hold
add 25 µL	ddbidest. H2O

Gel electrophoresis

Gel substances

1% Agarose gel
10 µl Ethidium bromide 50 ml gel
4 µl 2-log DNA ladder (New England Biolabs)

Lane

Content

Expectations

2-5

pSB1A3 iBB486476315 K1-4

~ 6,000 bp for pSB1A3_486476315

280 bp for BBa_K1071005

Each sample contained 2 bands, that means we got all expected bands and it could be assumed that the insertion of BBa_K1071005 was successful.

25.06.2013

Digest

Investigator: Dominik

Aim: Test digest of pSB1A3-iBB39, pSB1A3-iBB49 and pSB1C3-iBB6315 with EcoRI and PstI

1 µl Plasmid (pSB1C3-iBB39,pSB1A3-iBB49 and pSB1C3-iBB6315)
0.3 µl EcoRI
0.3 µl PstI
1 µl CutSmart
7.4 µl ddbidest. H2O

The samples were incubated for 1 h at 37 °C.

Worked as expected.

26.06.2013

Digest

Investigator: Dominik

Aim: Construction of the plasmids pSB1A3-iBB10+9, pSB1A3-iBB11+9, pSB1A3-iBB12+9 and pSB1A3-iBB13+9

1 µl Plasmid (pSB1C3-iBB10, pSB1C3-iBB11, pSB1C3-iBB12 and pSB1C3-iBB13)
0.5 µl EcoRI
0.5 µl SpeI
2 µl CutSmart
12 µl ddbidest. H2O

1 µl Plasmid (pSB1C3-iBB9)
0.5 µl XbaI
0.5 µl Pst/I
2 µl CutSmart
12 µl ddbidest. H2O

1 µl Plasmid (pSB1A3)
0.5 µl EcoRI
0.5 µl PstI
2 µl CutSmart
12 µl ddbidest. H2O

The samples were incubated for 1.5 h at 37 °C.

All expected fragments were present. The relevant fragments were cut from the gel and then purified via QIAgen Gel Extraction Kit (Qiagen).

Ligation

Investigator: Dominik

Aim: Assemble the biobricks iBB10+iBB9, iBB11+iBB9, iBB12+iBB9 and iBB13+iBB9 into the vector pSB1A3.

2 µl vector DNA (pSB1A3)
2 µl insert 1 DNA (iBB9)
2 µl insert 2 DNA (iBB10, iBB11, iBB12 or iBB13)
2 µl 10x T4 DNA ligase buffer
2 µl T4 DNA ligase
2 µl ddbidest. H2O

The samples were incubated overnight at room temperature.

27.06.2013

Transformation

Investigator: Dominik

Aim: Transformation of E. coli DH5α with the plasmid DNA pSB1A3-iBB10+iBB9, pSB1A3-iBB11+iBB9, pSB1A3-iBB12+iBB9 and pSB1A3-iBB13+iBB9.

The chemo competent E. coli DH5α cells were transformed with the plasmids pSB1A3-iBB10+iBB9, pSB1A3-iBB11+iBB9,pSB1A3-iBB12+iBB9 and pSB1A3-iBB13+iBB9 DNA and plated on LB-Amp-plates.
The plates were incubated over night at 37 °C.

Inoculation

Investigator: Dominik

Aim: inoculation of colonies for plasmid preparation

4 colonies of pSB1A3-iBB395146 and 4 colonies of pSB1A3-iBB496315 were inoculated in 5 ml LB medium containing ampicillin.

Digest

Investigator: Alex

Aim: Digest of pSB1A3-iBB395416 and pSB1A3-iBB496315 with EcoRI and PstI.

1 µl Plasmid DNA
0.3 µl EcoRI
0.3 µl PstI
1 µl Cut Smart
7.4 µl ddbidest. H2O

The samples were incubated for 2 h at 37 °C.

Gel electrophoresis

Gel substances

1% Agarose gel
10 µl Ethidium bromide in 50 ml gel
4 µl 2-log DNA ladder (New England Biolabs)

Lane	Content	Expectations
upper:
2-5	iBB395416 K1-4	2400 + 2000 bp
6	iBB39	1000 + 2000 bp
7	iBB6416	1400 + 2000 bp
lower:
2-6	iBB496315 K1-5	2400 + 2000 bp
7	iBB49	1200 + 2000 bp

All clones show a double band above 2000 bp.

We received all expected fragments. The success of the ligation should be double-checked by means of PCR.

@@ Line 1: / Line 1: @@
 {{:Team:Marburg/Template:Header}}
-{{:Team:Marburg/Template:ContentTitle}}
+{{:Team:Marburg/Template:ContentTitleNav}}
-Notebook: June
+Notebook: June <html><a href="https://2013.igem.org/Team:Marburg/Notebook:July"><img src="https://static.igem.org/mediawiki/2013/7/71/Mr-igem-next-arrow.png" style="float:right;margin-left:5px !important;" alt="Next"></a>&nbsp;<a href="https://2013.igem.org/Team:Marburg/Notebook:Mai"><img src="https://static.igem.org/mediawiki/2013/1/13/Mr-igem-previous-arrow.png" alt="Previous" style="float:right;"></a></html>
-{{:Team:Marburg/Template:ContentStart}}
+{{:Team:Marburg/Template:ContentStartNav}}<html>
-<html>
 <div class="notebooky-entry">
 <h2 class="title">
-	<a name="17-06-2013">17.06.2013</a>
+	<a name="03-06-2013">03.06.2013</a>
 </h2>
+<!-- Digest -->
 <fieldset class="experiment digest">
      <legend><a name="dig">Digest</a></legend>
      <div class="investigator">
 		<span class="inv">Investigator:</span>
-		<span class="inv-names">Alex</span>
+		<span class="inv-names">Franzi</span>
 	</div>
      <div class="aim">
 		<span class="aim">Aim:</span>
-		<span class="aim-desc">Digest of pSB1A3_3+9 and pSB1A3_4+9 with <i>Spe</i>I and <i>Pst</i>I, and corresponding inserts pSB1C3_5+4+1+6 and pSB1C3_6+3+1+5 with <i>EcoR</i>I and <i>Pst</i>I.</span>
+		<span class="aim-desc">Construction of P<sub>NR</sub>-test-vector &rarr; Digestion of combined 2 BioBricks.</span>
 	</div>
 	<div class="exp-content">
 		<ul class="digest">
- 			<li>1 µl DNA</li>
+			<li>10 µl DNA template</li>
-			<li>0.3 µl Enzyme 1 (SpeI or EcoRI, resp.)</li>
+			<li>0,5 µl enzyme 1</li>
-			<li>0.3 µl Enzyme 2 (PstI)</li>
+			<li>0,5 µl enzyme 2</li>
-			<li>1 µl Cut Smart</li>
+			<li>2 µl Cut Smart buffer</li>
-			<li>7.4 µl ddH2O</li>
+			<li>7 µl bidest. H2O</li>
 		</ul>
-		<p>The samples were incubated for 1.5 h at 37° C.</p>
+		<p>Samples:</p>
-                <p>Approaches were unsewed by means of gel electrophoresis, corresponding bands were picked under UV, DNA extraction from gel was performed using DNA Gel Extraction Kit (<i>QIAGEN</i>).</p>
+		<p>Digestion of pSB1A3 iBB1+5 with <i>Xba</i>I and <i>Pst</i>I-HF</p>
+		<p>Digestion of pSB1A3 iBB6+3 with <i>EcoR</i>I-HF and <i>Spe</i>I-HF.</p>
+		<p>The samples were incubated for 1 h at 37 °C.</p>
+		<p>They were purified via QIAgen Gel Extraction Kit (Qiagen). Elution into 20 µl bidest. H2O.</p>
+	</div>
 </fieldset>
+<!-- Ligation -->
 <fieldset class="experiment ligation">
-     <legend><a name="lig">Ligation, Transformation, Plasmid prep</a></legend>
+     <legend><a name="lig">Ligation</a></legend>
      <div class="investigator">
 		<span class="inv">Investigator:</span>
-		<span class="inv-names">Alex</span>
+		<span class="inv-names">Franzi</span>
 	</div>
      <div class="aim">
 		<span class="aim">Aim:</span>
-		<span class="aim-desc">Assemble the combined bricks 5416 and 6315 in their appropriate vectors pSB1A3_39 and pSB1A3_49, resp.</span>
+		<span class="aim-desc">Construction of P<sub>NR</sub>-test-vector &rarr; Ligation of two combined 2 BioBricks in pSB1C3.</span>
 	</div>
 	<div class="exp-content">
 		<ul class="lig">
-			<li>Insert und plasmid were transformed using a 1:2 ratio. Samples were incubated ON at RT.</li>
+			<li>45 ng vector DNA (pSB1C3; <i>EcoR</i>I/<i>Pst</i>I-cut)</li>
-			<li>Ligation product were transformed into chemical competent DH5α cells.</li>
+			<li>140 ng iBB6+3 (<i>EcoR</i>I/<i>Pst</i>I-cut)</li>
-			<li>Transformed cells were spread out on LB Amp culture plates.</li>
+			<li>150 ng iBB1+5 (<i>EcoR</i>I/<i>Pst</i>I-cut)</li>
-			<li>Incubation ON at RT.</li>
+			<li>2 µl 10x T4 DNA ligase buffer</li>
-                        <li>4 clones were picked for overnight cultures.</li>
+			<li>1 µl T4 DNA ligase</li>
-                        <li>A plasmid preparation was carried out the next day.</li>
+			<li>ad 20 µl bidest. H2O</li>
 		</ul>
+		<p>The samples were incubated for 1 h at RT.</p>
 	</div>
 </fieldset>
-</div>
-<div class="notebooky-entry">
+<!-- Transformation-->
-<h2 class="title">
+<fieldset class="experiment transformation">
-	<a name="22-06-2013">22.06.2013</a>
+    <legend>Transformation</legend>
-</h2>
+    <div class="investigator">
+		<span class="inv">Investigator:</span>
+		<span class="inv-names">Franzi</span>
+	</div>
+    <div class="aim">
+		<span class="aim">Aim:</span>
+		<span class="aim-desc">Construction of P<sub>NR</sub>-test-vector &rarr; Transformation of combined 4 BioBricks in pSB1C3 into <i>E. coli</i> DH5&alpha;.</span>
+	</div>
+	<div class="exp-content">
+		<p>Complete ligation samples were transformed into <i>E. coli</i> DH5&alpha; (30 min on ice, 60 sec 42 °C, 60 min 37 °C + 900 µl LB), plated on LB<sub>CM</sub>, overnight at 37 °C.</p>
+	</div>
+</fieldset>
-<fieldset class="experiment digest">
+<!-- Ligation -->
-     <legend><a name="dig">Test digest</a></legend>
+<fieldset class="experiment ligation">
+     <legend><a name="lig">Ligation</a></legend>
      <div class="investigator">
 		<span class="inv">Investigator:</span>
-		<span class="inv-names">Alex</span>
+		<span class="inv-names">Franzi</span>
 	</div>
      <div class="aim">
 		<span class="aim">Aim:</span>
-		<span class="aim-desc">Digest of pSB1A3_395416 and pSB1A3_496315 with <i>EcoR</i>I and <i>Pst</i>I.</span>
+		<span class="aim-desc">Construction of BioBricks &rarr; Ligation of single BioBricks into pSB1C3.</span>
 	</div>
 	<div class="exp-content">
-		<ul class="digest">
+		<ul class="lig">
-			<li>1 µl Plasmid DNA</li>
+			<li>45 ng vector DNA (pSB1C3; <i>EcoR</i>I/<i>Pst</i>I-cut)</li>
-			<li>0.3 µl EcoRI</li>
+			<li>X ng insert</li>
-			<li>0.3 µL PstI</li>
+			<li>1 µl 10x T4 DNA ligase buffer</li>
-			<li>1 µl Cut Smart</li>
+			<li>1 µl T4 DNA ligase</li>
-			<li>7.4 µl ddH2O</li>
+			<li>ad 10 µl bidest. H2O</li>
 		</ul>
-		<p>The samples were incubated for 2 h at 37° C.</p>
+		<p>Samples (all <i>EcoR</i>I/<i>Pst</i>I-cut):</p>
-		<table class="gel digest">
+			<p>120 ng eGFP (iBB9)</p>
-			<colgroup>
+				<p>60 ng SPTP1 (iBB10)</p>
-				<col width="50%" />
+				<p>50 ng SPTP2 (iBB11)</p>
-				<col width="50%" />
+				<p>55 ng SP1 (iBB12)</p>
-			</colgroup>
+				<p>50 ng SP2 (iBB13)</p>
-		<thead>
+		<p>The samples were incubated for 3 h at RT.</p>
-			<tr>
-				<th colspan="2" class="title">Gel electrophoresis</th>
-			</tr>
-		</thead>
-		<tbody>
-			<tr>
-				<td>
-					<img src="beispiel.png" width="50%" alt="gel-electrophoresis-image" />
-				</td>
-				<td>
-					<p>
-					<span class="gel-elc">Gel substances</span>
-					<ul class="gel-sub">
-						<li>1% Agarose gel</li>
-						<li>10 µl Ethidium bromide 50 ml gel</li>
-						<li>4 µl 2-log DNA ladder</li>
-					</ul>
-					</p>
-					<p>
-						<span class="exp">Expectations (upper gel)</span>
-						<ul class="exp">
-							<li>Lane 2: 1,200 + 2,000 bp</li>
-							<li>Lane 3: 1,200 + 2,000 bp</li>
-							<li>Lane 4-7: 2,400 + 2,000 bp</li>
-						</ul></p>
-                                                <p><span class="exp">Expectations (lower gel)</span>
-						<ul class="exp">
-							<li>Lane 2: 1,500 + 2,000 bp</li>
-							<li>Lane 3: 1,000 + 2,000 bp</li>
-							<li>Lane 4-7: 2,400 + 2,000 bp</li>
-						</ul></p>
-					</p>
-				</td>
-			</tr>
-			<tr>
-				<td colspan="2" class="gel-fazit">
-					<p>We didn’t receive all expected fragments. All expected fragments in lanes 4-7 were missing. Ligation failed :(</p>
-				</td>
-			</tr>
-		</tbody>
-		</table>
 	</div>
 </fieldset>
-</div>
+<!-- Transformation-->
+<fieldset class="experiment transformation">
+    <legend>Transformation</legend>
+    <div class="investigator">
+		<span class="inv">Investigator:</span>
+		<span class="inv-names">Franzi</span>
+	</div>
+    <div class="aim">
+		<span class="aim">Aim:</span>
+		<span class="aim-desc">Construction of BioBricks &rarr; Transformation of single BioBricks in pSB1C3 into <i>E. coli</i> DH5&alpha;.</span>
+	</div>
+	<div class="exp-content">
+		<p>Complete ligation samples were transformed into <i>E. coli</i> DH5&alpha; (30 min on ice, 60 sec 42 °C, 60 min 37 °C + 900 µl LB), plated on LB<sub>CM</sub>, overnight at 37 °C.</p>
+	</div>
+</fieldset>
-<div class="notebooky-entry">
+<!-- Inoculation -->
-<h2 class="title">
+<fieldset class="experiment inoculation">
-	<a name="24-06-2013">24.06.2013</a>
+     <legend><a name="ino">Inoculation</a></legend>
-</h2>
-<fieldset class="experiment pcr">
-     <legend><a name="pcr">PCR</a></legend>
      <div class="investigator">
 		<span class="inv">Investigator:</span>
-		<span class="inv-names">Alex</span>
+		<span class="inv-names">Franzi</span>
 	</div>
      <div class="aim">
 		<span class="aim">Aim:</span>
-		<span class="aim-desc">Check right assembly of biobrick BBa_K1071005 in the plasmid for antibody production</span>
+		<span class="aim-desc">Construction of P<sub>fcpB</sub>-test-vector and antibody-vector &rarr; Inoculation of transformants for miniprep.</span>
 	</div>
 	<div class="exp-content">
-		<table class="pcr">
+		<p>3 colonies of pSB1A3 iBB1+6 inoculated in 5 ml LB<sub>amp</sub>, 4 colonies of pSB1C3 iBB4864 inoculated in 5 ml LB<sub>CM</sub>, overnight at 37 °C.</p>
-			<colgroup>
-				<col width="15%" />
-				<col width="30%" />
-				<col width="5%" />
-				<col width="20%" />
-				<col width="20%" />
-				<col width="10%" />
-			</colgroup>
-			<thead>
-				<th>Volume</th>
-				<th>Reagent</th>
-				<th>&nbsp;</th>
-				<th>Temp (°C)</th>
-				<th colspan="2">Time</th>
-			</thead>
-			<tr>
-				<td>1 µl</td>
-				<td>pSB1A3_486476315</td>
-				<td>&nbsp;</td>
-				<td>95</td>
-				<td colspan="2">3 min</td>
-			</tr>
-			<tr>
-				<td>0.5 µl</td>
-				<td>Primer fwd</td>
-				<td>&nbsp;</td>
-				<td>95</td>
-				<td colspan="2">30 sec</td>
-			</tr>
-			<tr>
-				<td>0.5 µl</td>
-				<td>Primer rev</td>
-				<td>&nbsp;</td>
-				<td>60</td>
-				<td>30 sec</td>
-				<td rowspan="3">x30</td>
-			</tr>
-			<tr>
-				<td>0.5 µl</td>
-				<td>dNTPs</td>
-				<td>&nbsp;</td>
-				<td>72</td>
-				<td>42 sec</td>
-			</tr>
-			<tr>
-				<td>0.5 µl</td>
-				<td>Phusion polymerase</td>
-				<td>&nbsp;</td>
-				<td>72</td>
-				<td>5 min</td>
-			</tr>
-			<tr>
-				<td>5 µl</td>
-				<td>Phusion buffer 5x</td>
-				<td>&nbsp;</td>
-				<td>4</td>
-				<td colspan="2"><span class="hold">Hold</span></td>
-			</tr>
-                        <tr>
-				<td>add 25 µL</td>
-				<td>ddH2O</td>
-				<td>&nbsp;</td>
-			</tr>
-		</table>
-		<br />
-		<!-- Das Gel-Bild fast gleich wie beim Verdau -->
-		<table class="gel pcr">
-			<colgroup>
-				<col width="50%" />
-				<col width="50%" />
-			</colgroup>
-		<thead>
-			<tr>
-				<th colspan="2" class="title">Gel electrophoresis</th>
-			</tr>
-		</thead>
-		<tbody>
-			<tr>
-				<td>
-					<img src="beispiel.png" width="50%" alt="gel-electrophoresis-image" />
-				</td>
-				<td>
-					<p>
-					<span class="gel-elc">Gel substances</span>
-					<ul class="gel-sub">
-						<li>1% Agarose gel</li>
-						<li>10 µl Ethidium bromide 50 ml gel</li>
-						<li>4 µl 2-log DNA ladder</li>
-					</ul>
-					</p>
-					<p>
-						<span class="exp">Expactations</span>
-						<ul class="exp">
-							<li>Lane 1: ~ 6,000 bp for pSB1A3_486476315</li>
-							<li>Lane 2:     280 bp for BBa_K1071005</li>
-						</ul>
-					</p>
-					<p>Each sample contained 2 bands, that means we got all expected bands and it could be assumed that the insertion of BBa_K1071005 was successful.</p>
-				</td>
-			</tr>
-		</tbody>
-		</table>
 	</div>
 </fieldset>
 </div>
 <div class="notebooky-entry">
 <h2 class="title">
-	<a name="25-06-2013">25.06.2013</a>
+	<a name="04-06-2013">04.06.2013</a>
 </h2>
-<fieldset class="experiment digest">
+<!-- Miniprep -->
-     <legend><a name="dig">Digest</a></legend>
+<fieldset class="experiment miniprep">
+     <legend><a name="min">Miniprep</a></legend>
      <div class="investigator">
 		<span class="inv">Investigator:</span>
-		<span class="inv-names">Dominik</span>
+		<span class="inv-names">Franzi</span>
 	</div>
      <div class="aim">
 		<span class="aim">Aim:</span>
-		<span class="aim-desc">Test digest of pSB1A3-iBB39, pSB1A3-iBB49 and pSB1C3-iBB6315 with <i>Eco</i>RI and <i>Pst</i>I</span>
+		<span class="aim-desc">Construction of P<sub>fcpB</sub>-test-vector and antibody-vector &rarr; Miniprep of pSB1A3 iBB1+6 and pSB1C3 iBB4864.</span>
 	</div>
 	<div class="exp-content">
-		<p><ul class="digest">
+		<p>Miniprep of the liquid cultures (“QIAprep Spin Miniprep Kit” (Qiagen, Düsseldorf)), eluted in 30 µl bidest. H2O. 2 Cultures of pSB1C3 iBB4864 discarded (red colour).</p>
-			<li>1 µl Plasmid (pSB1C3-iBB39,pSB1A3-iBB49 and pSB1C3-iBB6315)</li>
-			<li>0.3 µl EcoRI</li>
-			<li>0.3 µl PstI</li>
-			<li>1 µl CutSmart</li>
-			<li>7.4 µl ddH2O</li>
-		</ul>
-		<p>The samples were incubated for 1 h at 37° C.</p>
-		<table class="gel digest">
-			<colgroup>
-				<col width="50%" />
-				<col width="50%" />
-			</colgroup>
-		<thead>
-			<tr>
-				<th colspan="2" class="title">Gel electrophoresis</th>
-			</tr>
-		</thead>
-		<tbody>
-			<tr>
-				<td>
-					<img src="beispiel.png" width="50%" alt="gel-electrophoresis-image" />
-				</td>
-				<td>
-					<p>
-					<span class="gel-elc">Gel substances</span>
-					<ul class="gel-sub">
-						<li>1% Agarose gel</li>
-						<li>10 µl RedSafe in 50 ml gel</li>
-						<li>x µl Hyper Ladder</li>
-					</ul>
-					</p>
-					<p>
-						<span class="exp">Expactations</span>
-						<ul class="exp">
-							<li>Lane 1: 5300 kbp</li>
-							<li>Lane 2: 3300 kbp</li>
-							<li>Lane 3: 1300 kbp</li>
-						</ul>
-					</p>
-					<p>Worked as expected.</p>
-				</td>
-			</tr>
-		</tbody>
-		</table>
 	</div>
 </fieldset>
-</div>
-<div class="notebooky-entry">
-<h2 class="title">
-	<a name="26-06-2013">26.06.2013</a>
-</h2>
+<!-- Digest -->
 <fieldset class="experiment digest">
      <legend><a name="dig">Digest</a></legend>
      <div class="investigator">
 		<span class="inv">Investigator:</span>
-		<span class="inv-names">Dominik</span>
+		<span class="inv-names">Franzi</span>
 	</div>
      <div class="aim">
 		<span class="aim">Aim:</span>
-		<span class="aim-desc">Construction of the plasmids pSB1A3-iBB10+9, pSB1A3-iBB11+9, pSB1A3-iBB12+9 and pSB1A3-iBB13+9</span>
+		<span class="aim-desc">Construction of P<sub>fcpB</sub>-test-vector and antibody-vector &rarr; Control digestion of pSB1A3 iBB1+6 and pSB1C3 iBB4864.</span>
 	</div>
 	<div class="exp-content">
-		<p><ul class="digest">
+		<ul class="digest">
-			<li>1 µl Plasmid (pSB1C3-iBB10, pSB1C3-iBB11, pSB1C3-iBB12 and pSB1C3-iBB13)</li>
+			<li>1 µl DNA template</li>
-			<li>0.5 µl EcoRI</li>
+			<li>0,5 µl <i>EcoR</i>I-HF</li>
-			<li>0.5 µl SpeI</li>
+			<li>0,5 µl <i>Pst</i>I-HF</li>
-			<li>2 µl CutSmart</li>
+			<li>1 µl Cut Smart buffer</li>
-			<li>12 µl ddH2O</li>
+			<li>7 µl bidest. H2O</li>
 		</ul>
+		<p>The samples were incubated for 2 h at 37 °C.</p>
-		<p><ul class="digest">
-			<li>1 µl Plasmid (pSB1C3-iBB9)</li>
-			<li>0.5 µl XbaI</li>
-			<li>0.5 µl PstI</li>
-			<li>2 µl CutSmart</li>
-			<li>12 µl ddH2O</li>
-		</ul>
-		<p><ul class="digest">
-			<li>1 µl Plasmid (pSB1A3)</li>
-			<li>0.5 µl EcoRI</li>
-			<li>0.5 µl PstI</li>
-			<li>2 µl CutSmart</li>
-			<li>12 µl ddH2O</li>
-		</ul>
-		<p>The samples were incubated for 1.5 h at 37° C.</p>
 		<table class="gel digest">
 			<colgroup>
@@ Line 376: / Line 193: @@
 			<tr>
 				<td>
-					<img src="beispiel.png" width="50%" alt="gel-electrophoresis-image" />
+					<img src="https://static.igem.org/mediawiki/2013/5/5f/Gel_2013-06-04.png" width="100%" alt="gel-electrophoresis-image" />
 				</td>
 				<td>
@@ Line 383: / Line 200: @@
 					<ul class="gel-sub">
 						<li>1% Agarose gel</li>
-						<li>10 µl RedSafe in 50 ml gel</li>
+						<li>10 µl RedSafe per 50 ml gel</li>
-						<li>x µl Hyper Ladder</li>
+						<li>6 µl 2-Log DNA Ladder (NEB)</li>
+						<li>6x loading buffer (for samples)</li>
 					</ul>
 					</p>
 					<p>
-						<span class="exp">Expactations</span>
+							<table class="gel">
-						<ul class="exp">
+								<colgroup>
-							<li>Lane 1: 5300 kbp</li>
+									<col width="10%" />
-							<li>Lane 2: 3300 kbp</li>
+									<col width="2%" />
-							<li>Lane 3: 1300 kbp</li>
+									<col width="50%" />
-						</ul>
+									<col width="5%" />
+									<col width="33%" />
+								</colgroup>
+								<thead>
+									<th>Lane</th>
+									<th>&nbsp;</th>
+									<th>Content</th>
+									<th>&nbsp;</th>
+									<th>Expectations</th>
+								</thead>
+								<tr>
+									<td>2+3</td>
+									<th>&nbsp;</th>
+									<td>iBB4864 K1-2</td>
+									<th>&nbsp;</th>
+									<td>2580 bp</td>
+								</tr>
+								<tr>
+									<td>4</td>
+									<th>&nbsp;</th>
+									<td>empty</td>
+									<th>&nbsp;</th>
+									<td></td>
+								<tr>
+									<td>5-7</td>
+									<th>&nbsp;</th>
+									<td>iBB16 K3-5</td>
+									<th>&nbsp;</th>
+									<td>650 bp</td>
+								</tr>
+							</table>
 					</p>
-					<p>All expected fragments were present. The relevant fragments were cut from the gel and then purified via “Qiagen Gel Extraction Kit” (Qiagen, Düsseldorf).</p>
 				</td>
 			</tr>
@@ Line 403: / Line 250: @@
 </fieldset>
-<fieldset class="experiment ligation">
+<!-- Digest -->
-     <legend><a name="lig">Ligation</a></legend>
+<fieldset class="experiment digest">
+     <legend><a name="dig">Digest</a></legend>
      <div class="investigator">
 		<span class="inv">Investigator:</span>
-		<span class="inv-names">Dominik</span>
+		<span class="inv-names">Franzi</span>
 	</div>
      <div class="aim">
 		<span class="aim">Aim:</span>
-		<span class="aim-desc">Assemble the biobricks iBB10+iBB9, iBB11+iBB9, iBB12+iBB9 and iBB13+iBB9 into the vector pSB1A3.</span>
+		<span class="aim-desc">Construction of P<sub>fcpB</sub>-test-vector and antibody-vector &rarr; Digestion of combined 2 BioBricks in pSB1A3 and combined 4 BioBricks in pSB1C3.</span>
 	</div>
 	<div class="exp-content">
-		<p><ul class="lig">
+		<ul class="digest">
-			<li>2 µl vector DNA (pSB1A3)</li>
+			<li>10 µl DNA template</li>
-			<li>2 µl insert 1 DNA (iBB9)</li>
+			<li>0,5 µl enzyme 1</li>
-			<li>2 µl insert 2 DNA (iBB10, iBB11, iBB12 or iBB13)</li>
+			<li>0,5 µl enzyme 2</li>
-			<li>2 µl 10x T4 DNA ligase buffer</li>
+			<li>2 µl Cut Smart buffer</li>
-			<li>2 µl T4 DNA ligase</li>
+			<li>7 µl bidest. H2O</li>
-			<li>2 µl ddH2O</li>
 		</ul>
-		<p>The samples were incubated overnight at room temperature.</p>
+		<p>Samples:</p>
+		<p>Digestion of pSB1C3 iBB7631 K4 and pSB1A3 iBB16 K4 with <i>Xba</i>I and <i>Pst</i>I-HF</p>
+		<p>Digestion of pSB1C3 iBB4864 K1 and pSB1A3 iBB54 K2 with <i>EcoR</i>I-HF and <i>Spe</i>I-HF.</p>
+		<p>The samples were incubated for 1 h at 37 °C.</p>
+		<p>They were purified via QIAgen Gel Extraction Kit (Qiagen). Elution into 20 µl bidest. H2O.</p>
 	</div>
 </fieldset>
-</div>
-<div class="notebooky-entry">
+<!-- Ligation -->
-<h2 class="title">
+<fieldset class="experiment ligation">
-	<a name="27-06-2013">27.06.2013</a>
+    <legend><a name="lig">Ligation</a></legend>
-</h2>
-<fieldset class="experiment transformation">
-    <legend>Transformation</legend>
      <div class="investigator">
 		<span class="inv">Investigator:</span>
-		<span class="inv-names">Dominik</span>
+		<span class="inv-names">Franzi</span>
 	</div>
      <div class="aim">
 		<span class="aim">Aim:</span>
-		<span class="aim-desc">Transformation of <i>E. coli</i> DH5&#945; with the plasmid DNA pSB1A3-iBB10+iBB9, pSB1A3-iBB11+iBB9, pSB1A3-iBB12+iBB9 and pSB1A3-iBB13+iBB9.</span>
+		<span class="aim-desc">Construction of P<sub>fcpB</sub>-test-vector and antibody-vector &rarr; Ligation of two combined 2 BioBricks into pSB1C3 and two combined 4 BioBricks into pSB1A3.</span>
 	</div>
 	<div class="exp-content">
-		<p>The chemo competent <i>E. coli</i> DH5&alpha; cells were transformed with the plasmids pSB1A3-iBB10+iBB9, pSB1A3-iBB11+iBB9,pSB1A3-iBB12+iBB9 and pSB1A3-iBB13+iBB9 DNA and plated on LB-Amp-plates.<br />
+	<p>Antibody:</p>
-		The plates were incubated over night at 37° C.</p>
+		<ul class="lig">
+			<li>80 ng vector DNA (pSB1A3; <i>EcoR</i>I/<i>Pst</i>I-cut)</li>
+			<li>160 ng iBB7631</li>
+			<li>190 ng iBB4864 </li>
+			<li>1 µl 10x T4 DNA ligase buffer</li>
+			<li>1 µl T4 DNA ligase</li>
+			<li>ad 10 µl bidest. H2O</li>
+		</ul>
+	<p>Test-vector:</p>
+		<ul class="lig">
+			<li>45 ng vector DNA (pSB1C3; <i>EcoR</i>I/<i>Pst</i>I-cut)</li>
+			<li>150 ng iBB54</li>
+			<li>120 ng iBB16</li>
+			<li>1 µl 10x T4 DNA ligase buffer</li>
+			<li>1 µl T4 DNA ligase</li>
+			<li>ad 10 µl bidest. H2O</li>
+		</ul>
+		<p>Both samples were incubated overnight at 16 °C.</p>
 	</div>
 </fieldset>
+<!-- Inoculation -->
 <fieldset class="experiment inoculation">
      <legend><a name="ino">Inoculation</a></legend>
      <div class="investigator">
 		<span class="inv">Investigator:</span>
-		<span class="inv-names">Dominik</span>
+		<span class="inv-names">Franzi</span>
 	</div>
      <div class="aim">
 		<span class="aim">Aim:</span>
-		<span class="aim-desc">inoculation of colonies for plasmid preparation</span>
+		<span class="aim-desc">Construction of P<sub>NR</sub>-test-vector and BioBricks &rarr; Inoculation of transformants.</span>
 	</div>
 	<div class="exp-content">
-		<p>4 colonies of pSB1A3-iBB395146 and 4 colonies of pSB1A3-iBB496315 were inoculated in 5 ml LB medium containing ampicillin.</p>
+		<p>Transformations (03.06.13) inoculated in 5 ml LB<sub>CM</sub>, overnight at 37 °C.</p>
 	</div>
 </fieldset>
 </div>
 <div class="notebooky-entry">
 <h2 class="title">
-	<a name="28-06-2013">28.06.2013</a>
+	<a name="05-06-2013">05.06.2013</a>
 </h2>
+<!-- Miniprep -->
+<fieldset class="experiment miniprep">
+    <legend><a name="min">Miniprep</a></legend>
+    <div class="investigator">
+		<span class="inv">Investigator:</span>
+		<span class="inv-names">Franzi</span>
+	</div>
+    <div class="aim">
+		<span class="aim">Aim:</span>
+		<span class="aim-desc">Construction of P<sub>NR</sub>-test-vector and BioBricks &rarr; Miniprep of single BioBricks and pSB1C3 iBB6315.</span>
+	</div>
+	<div class="exp-content">
+		<p>Miniprep of the liquid cultures (“QIAprep Spin Miniprep Kit” (Qiagen, Düsseldorf)), eluted in 30 µl bidest. H2O.</p>
+	</div>
+</fieldset>
+<!-- Digest -->
 <fieldset class="experiment digest">
      <legend><a name="dig">Digest</a></legend>
      <div class="investigator">
 		<span class="inv">Investigator:</span>
-		<span class="inv-names">Alex</span>
+		<span class="inv-names">Franzi</span>
 	</div>
      <div class="aim">
 		<span class="aim">Aim:</span>
-		<span class="aim-desc">Digest of pSB1A3-iBB395416 and pSB1A3-iBB496315 with <i>EcoR</i>I and <i>Pst</i>I.</span>
+		<span class="aim-desc">Construction of P<sub>NR</sub>-test-vector and BioBricks &rarr; Control digestion of single BioBricks and pSB1C3 iBB6315.</span>
 	</div>
 	<div class="exp-content">
 		<ul class="digest">
-			<li>1 µl Plasmid DNA</li>
+			<li>1 µl DNA template</li>
-			<li>0.3 µl EcoRI</li>
+			<li>0,3 µl <i>EcoR</i>I-HF</li>
-			<li>0.3 µl PstI</li>
+			<li>0,3 µl <i>Pst</i>I-HF</li>
-			<li>1 µl Cut Smart</li>
+			<li>1 µl Cut Smart buffer</li>
-			<li>7.4 µl ddH2O</li>
+			<li>7,4 µl bidest. H2O</li>
 		</ul>
-		<p>The samples were incubated for 2 h at 37° C.</p>
+		<p>The samples were incubated for 1,5 h at 37 °C.</p>
 		<table class="gel digest">
 			<colgroup>
@@ Line 501: / Line 384: @@
 			<tr>
 				<td>
-					<img src="beispiel.png" width="50%" alt="gel-electrophoresis-image" />
+					<img src="https://static.igem.org/mediawiki/2013/f/fd/Gel_2013-06-05_1.png" width="100%" alt="gel-electrophoresis-image" />
 				</td>
 				<td>
@@ Line 508: / Line 391: @@
 					<ul class="gel-sub">
 						<li>1% Agarose gel</li>
-						<li>10 µl Ethidium bromide in 50 ml gel</li>
+						<li>10 µl RedSafe per 50 ml gel</li>
-						<li>4 µl 2-log DNA ladder</li>
+						<li>6 µl 2-Log DNA Ladder (NEB)</li>
-					</ul>
+						<li>6x loading buffer (for samples)</li>
+						</ul>
 					</p>
 					<p>
-						<span class="exp">Expectations (upper gel)</span>
+							<table class="gel">
-						<ul class="exp">
+								<colgroup>
-							<li>Lane 2-5: 2,400 + 2,000 bp</li>
+									<col width="10%" />
-							<li>Lane 6: 1,000 + 2,000 bp</li>
+									<col width="2%" />
-							<li>Lane 7: 1,400 + 2,000 bp</li>
+									<col width="50%" />
-						</ul></p>
+									<col width="5%" />
-                                                <p><span class="exp">Expectations (lower gel)</span>
+									<col width="33%" />
-						<ul class="exp">
+								</colgroup>
-							<li>Lane 2-7: 2,400 + 2,000 bp</li>
+								<thead>
-							<li>Lane 8: 1,200 + 2,000 bp</li>
+									<th>Lane</th>
-						</ul></p>
+									<th>&nbsp;</th>
+									<th>Content</th>
+									<th>&nbsp;</th>
+									<th>Expectations</th>
+								</thead>
+								<tr>
+									<td>2-4</td>
+									<th>&nbsp;</th>
+									<td>iBB9 K1-3</td>
+									<th>&nbsp;</th>
+									<td>780 bp</td>
+								</tr>
+								<tr>
+									<td>5</td>
+									<th>&nbsp;</th>
+									<td>empty</td>
+									<th>&nbsp;</th>
+									<td></td>
+								</tr>
+								<tr>
+									<td>6-8</td>
+									<th>&nbsp;</th>
+									<td>iBB10 K1-3</td>
+									<th>&nbsp;</th>
+									<td>228 bp</td>
+								</tr>
+								<tr>
+									<td></td>
+									<th>&nbsp;</th>
+									<td></td>
+									<th>&nbsp;</th>
+									<td>pSB1C3: 2 kb</td>
+								</tr>
+							</table>
 					</p>
-					<p>All clones show a double band above 2,000 bp.</p>
 				</td>
 			</tr>
 			<tr>
-				<td colspan="2" class="gel-fazit">
+				<td>
-					<p>We received all expected fragments. The success of the ligation should be double-checked by means of PCR.</p>
+					<img src="https://static.igem.org/mediawiki/2013/f/f2/Gel_2013-06-05_2.png" width="100%" alt="gel-electrophoresis-image" />
+				</td>
+				<td>
+					<p>
+							<table class="gel">
+								<colgroup>
+									<col width="10%" />
+									<col width="2%" />
+									<col width="50%" />
+									<col width="5%" />
+									<col width="33%" />
+								</colgroup>
+								<thead>
+									<th>Lane</th>
+									<th>&nbsp;</th>
+									<th>Content</th>
+									<th>&nbsp;</th>
+									<th>Expectations</th>
+								</thead>
+								<tr>
+									<td>2-4</td>
+									<th>&nbsp;</th>
+									<td>iBB11 K1-3</td>
+									<th>&nbsp;</th>
+									<td>147 bp</td>
+								</tr>
+								<tr>
+									<td>5</td>
+									<th>&nbsp;</th>
+									<td>empty</td>
+									<th>&nbsp;</th>
+									<td></td>
+								</tr>
+								<tr>
+									<td>6-8</td>
+									<th>&nbsp;</th>
+									<td>iBB12 K1-3</td>
+									<th>&nbsp;</th>
+									<td>123 bp</td>
+								</tr>
+								<tr>
+									<td></td>
+									<th>&nbsp;</th>
+									<td></td>
+									<th>&nbsp;</th>
+									<td>pSB1C3: 2 kb</td>
+								</tr>
+							</table>
+					</p>
+				</td>
+			</tr>
+			<tr>
+				<td>
+					<img src="https://static.igem.org/mediawiki/2013/5/5a/Gel_2013-06-05_3.png" width="100%" alt="gel-electrophoresis-image" />
+				</td>
+				<td>
+					<p>
+							<table class="gel">
+								<colgroup>
+									<col width="10%" />
+									<col width="2%" />
+									<col width="50%" />
+									<col width="5%" />
+									<col width="33%" />
+								</colgroup>
+								<thead>
+									<th>Lane</th>
+									<th>&nbsp;</th>
+									<th>Content</th>
+									<th>&nbsp;</th>
+									<th>Expectations</th>
+								</thead>
+								<tr>
+									<td>2-4</td>
+									<th>&nbsp;</th>
+									<td>iBB13 K1-3</td>
+									<th>&nbsp;</th>
+									<td>108 bp</td>
+								</tr>
+								<tr>
+									<td>5</td>
+									<th>&nbsp;</th>
+									<td>empty</td>
+									<th>&nbsp;</th>
+									<td></td>
+								</tr>
+								<tr>
+									<td>6-8</td>
+									<th>&nbsp;</th>
+									<td>iBB6315 K1-3</td>
+									<th>&nbsp;</th>
+									<td>1200 bp</td>
+								</tr>
+								<tr>
+									<td></td>
+									<th>&nbsp;</th>
+									<td></td>
+									<th>&nbsp;</th>
+									<td>pSB1C3: 2 kb</td>
+								</tr>
+							</table>
+					</p>
 				</td>
 			</tr>
@@ Line 538: / Line 555: @@
 </fieldset>
-<div class="notebooky-entry">
+<!-- Transformation-->
-<h2 class="title">
+<fieldset class="experiment transformation">
-	<a name="02-07-2013">02.07.2013</a>
+     <legend>Transformation</legend>
-</h2>
-<fieldset class="experiment miniprep">
-     <legend><a name="min">Miniprep</a></legend>
      <div class="investigator">
 		<span class="inv">Investigator:</span>
-		<span class="inv-names">Dominik</span>
+		<span class="inv-names">Franzi</span>
 	</div>
      <div class="aim">
 		<span class="aim">Aim:</span>
-		<span class="aim-desc">preparation of the plasmids pSB1A3-iBB10+iBB9, pSB1A3-iBB11+iBB9, pSB1A3-iBB12+iBB9 and pSB1A3-iBB13+iBB9.</span>
+		<span class="aim-desc">Construction of P<sub>fcpB</sub>-test-vector and antibody-vector &rarr; Transformation of combined 4 BioBricks in pSB1C3 and combined 8 BioBricks in pSB1A3 into <i>E. coli</i> DH5&alpha;.</span>
 	</div>
 	<div class="exp-content">
-		<p>The plasmids were isolated with the “QIAprep Spin Miniprep Kit” (Qiagen, Düsseldorf) according to the producer instructions.</p>
+		<p>Ligations (04.06.13) were transformed into <i>E. coli</i> DH5&alpha; (30 min on ice, 60 sec 42 °C, 60 min (1C3)/40 min (1A3) 37 °C + 900 µl LB), plated on LB<sub>CM</sub> (1C3)/LB<sub>amp</sub> (1A3), overnight at 37 °C.</p>
 	</div>
 </fieldset>
-<fieldset class="experiment digest">
+<!-- Sequencing -->
-     <legend><a name="dig">Digest</a></legend>
+<fieldset class="experiment sequencing">
+     <legend>Sequencing</legend>
      <div class="investigator">
 		<span class="inv">Investigator:</span>
-		<span class="inv-names">Dominik</span>
+		<span class="inv-names">Franzi</span>
 	</div>
      <div class="aim">
 		<span class="aim">Aim:</span>
-		<span class="aim-desc">Test digest of the plasmids pSB1A3-iBB10+iBB9, pSB1A3-iBB11+iBB9, pSB1A3-iBB12+iBB9 and pSB1A3-iBB13+iBB9 with <i>Eco</i>RI and <i>Pst</i>I</span>
+		<span class="aim-desc">Construction of BioBricks &rarr; Examination of sequencing results of various constructs.</span>
 	</div>
 	<div class="exp-content">
-		<p><ul class="digest">
+		<p>iBB9 K1 fw - AGB000K 263</p>
-			<li>1 µl Plasmid (pSB1A3-iBB10+iBB9, pSB1A3-iBB11+iBB9, pSB1A3-iBB12+iBB9 and pSB1A3-iBB13+iBB9)</li>
+		<p>iBB9 K1 rv - AGB000K 264</p>
-			<li>0.3 µl EcoRI</li>
+		<p>iBB9 K2 fw - AGB000K 265</p>
-			<li>0.3 µl PstI</li>
+		<p>iBB9 K2 rv - AGB000K 266</p>
-			<li>1 µl CutSmart</li>
+		<p>iBB10 K1 fw - AGB000K 267</p>
-			<li>7.4 µl ddH2O</li>
+		<p>iBB10 K2 fw - AGB000K 268</p>
-		</ul>
+		<p>iBB11 K1 fw - AGB000K 269</p>
-		<p>The samples were incubated for 1 h at 37° C.</p>
+		<p>iBB11 K2 fw - AGB000K 270</p>
-		<table class="gel digest">
+		<p>iBB12 K1 fw - AGB000K 271</p>
-			<colgroup>
+		<p>iBB12 K2 fw - AGB000K 272</p>
-				<col width="50%" />
+		<p>iBB13 K1 fw - AGB000K 273</p>
-				<col width="50%" />
+		<p>iBB13 K2 fw - AGB000K 274</p>
-			</colgroup>
+		<p> </p>
-		<thead>
+		<p>all correct, except iBB10 G&rarr;T pos.30, iBB11 T&rarr;C pos.24. Probably template mutated.</p>
-			<tr>
-				<th colspan="2" class="title">Gel electrophoresis</th>
-			</tr>
-		</thead>
-		<tbody>
-			<tr>
-				<td>
-					<img src="beispiel.png" width="50%" alt="gel-electrophoresis-image" />
-				</td>
-				<td>
-					<p>
-					<span class="gel-elc">Gel substances</span>
-					<ul class="gel-sub">
-						<li>1% Agarose gel</li>
-						<li>10 µl RedSafe in 50 ml gel</li>
-						<li>x µl Hyper Ladder</li>
-					</ul>
-					</p>
-					<p>
-						<span class="exp">Expactations</span>
-						<ul class="exp">
-							<li>Lane 1: 5300 kbp</li>
-							<li>Lane 2: 3300 kbp</li>
-							<li>Lane 3: 1300 kbp</li>
-						</ul>
-					</p>
-					<p>Worked as expected.</p>
-				</td>
-			</tr>
-		</tbody>
-		</table>
 	</div>
 </fieldset>
 </div>
 <div class="notebooky-entry">
 <h2 class="title">
-	<a name="23-07-2013">23.07.2013</a>
+	<a name="06-06-2013">06.06.2013</a>
 </h2>
-<fieldset class="experiment digest">
+<!-- Inoculation -->
-     <legend><a name="dig">Digest</a></legend>
+<fieldset class="experiment inoculation">
+     <legend><a name="ino">Inoculation</a></legend>
      <div class="investigator">
 		<span class="inv">Investigator:</span>
-		<span class="inv-names">Dominik</span>
+		<span class="inv-names">Franzi</span>
 	</div>
      <div class="aim">
 		<span class="aim">Aim:</span>
-		<span class="aim-desc">Test digest of the plasmids pSB1C3-iBB4+iBB11+iBB9, pSB1C3-iBB4+iBB12+iBB9 with <i>Eco</i>RI and <i>Pst</i>I</span>
+		<span class="aim-desc">Construction of P<sub>fcpB</sub>-test-vector and antibody-vector &rarr; Inoculation of pSB1C3 iBB5416 and pSB1A3 iBB48647631 for miniprep.</span>
 	</div>
 	<div class="exp-content">
-		<p><ul class="digest">
+		<p>3 colonies of pSB1C3 iBB5416 inoculated in 5 ml LB<sub>CM</sub>, 4 colonies of pSB1A3 iBB48647631 inoculated in 5 ml LB<sub>amp</sub>, overnight at 37 °C.</p>
-			<li>1 µl Plasmid (pSB1C3-iBB4+iBB11+iBB9 and pSB1C3-iBB4+iBB12+iBB9)</li>
-			<li>0.3 µl EcoRI</li>
-			<li>0.3 µl PstI</li>
-			<li>1 µl CutSmart</li>
-			<li>7.4 µl ddH2O</li>
-		</ul>
-		<p>The samples were incubated for 1 h at 37° C.</p>
-		<table class="gel digest">
-			<colgroup>
-				<col width="50%" />
-				<col width="50%" />
-			</colgroup>
-		<thead>
-			<tr>
-				<th colspan="2" class="title">Gel electrophoresis</th>
-			</tr>
-		</thead>
-		<tbody>
-			<tr>
-				<td>
-					<img src="beispiel.png" width="50%" alt="gel-electrophoresis-image" />
-				</td>
-				<td>
-					<p>
-					<span class="gel-elc">Gel substances</span>
-					<ul class="gel-sub">
-						<li>1% Agarose gel</li>
-						<li>10 µl RedSafe in 50 ml gel</li>
-						<li>x µl Hyper Ladder</li>
-					</ul>
-					</p>
-					<p>
-						<span class="exp">Expactations</span>
-						<ul class="exp">
-							<li>Lane 1: 5300 kbp</li>
-							<li>Lane 2: 3300 kbp</li>
-							<li>Lane 3: 1300 kbp</li>
-						</ul>
-					</p>
-					<p>Worked as expected.</p>
-				</td>
-			</tr>
-		</tbody>
-		</table>
 	</div>
 </fieldset>
 </div>
 <div class="notebooky-entry">
 <h2 class="title">
-	<a name="24-07-2013">24.07.2013</a>
+	<a name="07-06-2013">07.06.2013</a>
 </h2>
-<fieldset class="experiment inoculation">
+<!-- Miniprep -->
-     <legend><a name="ino">Inoculation</a></legend>
+<fieldset class="experiment miniprep">
+     <legend><a name="min">Miniprep</a></legend>
      <div class="investigator">
 		<span class="inv">Investigator:</span>
-		<span class="inv-names">Dominik</span>
+		<span class="inv-names">Patrick</span>
 	</div>
      <div class="aim">
 		<span class="aim">Aim:</span>
-		<span class="aim-desc">inoculation of colonies for plasmid preparation</span>
+		<span class="aim-desc">Construction of P<sub>fcpB</sub>-test-vector and antibody-vector &rarr; Miniprep of pSB1C3 iBB5416 and pSB1A3 iBB48647631.</span>
 	</div>
 	<div class="exp-content">
-		<p>4 colonies of pSB1C3-iBB4+iBB10+iBB9 and 4 colonies of pSB1C3-iBB4+iBB13+iBB9 were inoculated in 5 ml LB medium containing chloramphenicol.</p>
+		<p>Miniprep of the liquid cultures (“QIAprep Spin Miniprep Kit” (Qiagen, Düsseldorf)), eluted in 30 µl bidest. H2O.</p>
 	</div>
 </fieldset>
+<!-- Digest -->
 <fieldset class="experiment digest">
      <legend><a name="dig">Digest</a></legend>
      <div class="investigator">
 		<span class="inv">Investigator:</span>
-		<span class="inv-names">Dominik</span>
+		<span class="inv-names">Patrick</span>
 	</div>
      <div class="aim">
 		<span class="aim">Aim:</span>
-		<span class="aim-desc">Digest of pSB1C3-iBB4+iBB11+iBB9, pSB1C3-iBB4+iBB12+iBB9, pSB1C3-iBB6315 and pSB1A3.</span>
+		<span class="aim-desc">Construction of P<sub>fcpB</sub>-test-vector and antibody-vector &rarr; Control digestion of pSB1C3 iBB5416 and pSB1A3 iBB48647631.</span>
 	</div>
 	<div class="exp-content">
-		<p><ul class="digest">
+		<ul class="digest">
-			<li>1000 ng Plasmid (pSB1C3-iBB4+iBB11+iBB9, pSB1C3-iBB4+iBB12+iBB9)</li>
+			<li>1 µl DNA template</li>
-			<li>0.5 µl EcoRI</li>
+			<li>0,3 µl <i>EcoR</i>I-HF</li>
-			<li>0.5 µl SpeI</li>
+			<li>0,3 µl <i>Pst</i>I-HF</li>
-			<li>2 µl CutSmart</li>
+			<li>1 µl Cut Smart buffer</li>
-			<li>ad 20 µl ddH2O</li>
+			<li>7,4 µl bidest. H2O</li>
 		</ul>
+		<p>The samples were incubated for 1 h at 37 °C.</p>
-		<p><ul class="digest">
-			<li>1000 ng Plasmid (pSB1C3-iBB6315)</li>
-			<li>0.5 µl XbaI</li>
-			<li>0.5 µl PstI</li>
-			<li>2 µl CutSmart</li>
-			<li>ad 20 µl ddH2O</li>
-		</ul>
-		<p><ul class="digest">
-			<li>1000 ng Plasmid (pSB1A3)</li>
-			<li>0.5 µl EcoRI</li>
-			<li>0.5 µl PstI</li>
-			<li>2 µl CutSmart</li>
-			<li>ad 20 µl ddH2O</li>
-		</ul>
-		<p>The samples were incubated for 1 h at 37° C.</p>
 		<table class="gel digest">
 			<colgroup>
@@ Line 752: / Line 681: @@
 			<tr>
 				<td>
-					<img src="beispiel.png" width="50%" alt="gel-electrophoresis-image" />
+					<img src="https://static.igem.org/mediawiki/2013/8/8b/Gel_2013-06-07.png" width="100%" alt="gel-electrophoresis-image" />
 				</td>
 				<td>
@@ Line 759: / Line 688: @@
 					<ul class="gel-sub">
 						<li>1% Agarose gel</li>
-						<li>10 µl RedSafe in 50 ml gel</li>
+						<li>10 µl RedSafe per 50 ml gel</li>
-						<li>x µl Hyper Ladder</li>
+						<li>6 µl 2-Log DNA Ladder (NEB)</li>
+						<li>6x loading buffer (for samples)</li>
 					</ul>
 					</p>
 					<p>
-						<span class="exp">Expactations</span>
+							<table class="gel">
-						<ul class="exp">
+								<colgroup>
-							<li>Lane 1: 5300 kbp</li>
+									<col width="10%" />
-							<li>Lane 2: 3300 kbp</li>
+									<col width="2%" />
-							<li>Lane 3: 1300 kbp</li>
+									<col width="50%" />
-						</ul>
+									<col width="5%" />
+									<col width="33%" />
+								</colgroup>
+								<thead>
+									<th>Lane</th>
+									<th>&nbsp;</th>
+									<th>Content</th>
+									<th>&nbsp;</th>
+									<th>Expectations</th>
+								</thead>
+								<tr>
+									<td>2-4</td>
+									<th>&nbsp;</th>
+									<td>iBB5416 K1-3</td>
+									<th>&nbsp;</th>
+									<td>
+										<p>pSB1C3: 2070 bp</p>
+										<p>iBB5416: 1500 bp</p>
+									</td>
+								</tr>
+								<tr>
+									<td>5-8</td>
+									<th>&nbsp;</th>
+									<td>iBB48647631 K1-4</td>
+									<th>&nbsp;</th>
+									<td>
+										<p>pSB1A3: 2150 bp</p>
+										<p>iBB48647631: 4157 bp</p>
+									</td>
+								</tr>
+							</table>
 					</p>
-					<p>All expected fragments were present. The relevant fragments were cut from the gel and then purified via “Qiagen Gel Extraction Kit” (Qiagen, Düsseldorf).</p>
 				</td>
 			</tr>
@@ Line 779: / Line 738: @@
 </fieldset>
-<fieldset class="experiment ligation">
+</div>
-     <legend><a name="lig">Ligation</a></legend>
+<div class="notebooky-entry">
+<h2 class="title">
+	<a name="10-06-2013">10.06.2013</a>
+</h2>
+<!-- Digest -->
+<fieldset class="experiment digest">
+     <legend><a name="dig">Digest</a></legend>
      <div class="investigator">
 		<span class="inv">Investigator:</span>
-		<span class="inv-names">Dominik</span>
+		<span class="inv-names">Lukas</span>
 	</div>
      <div class="aim">
 		<span class="aim">Aim:</span>
-		<span class="aim-desc">Assemble the biobricks iBB4+iBB11+iBB96315 and iBB4+iBB12+iBB96315 into the vector pSB1A3.</span>
+		<span class="aim-desc">Construction of P<sub>NR</sub>-test-vector, P<sub>fcpB</sub>-test-vector and antibody-vector &rarr; Digestion of pSB1A3 iBB48647631 and single BioBricks.</span>
 	</div>
 	<div class="exp-content">
-		<p><ul class="lig">
+		<ul class="digest">
-			<li>2 µl vector DNA (pSB1A3)</li>
+			<li>X µl DNA template</li>
-			<li>5 µl insert 1 DNA (iBB6315)</li>
+			<li>0,5 µl enzyme 1</li>
-			<li>5 µl insert 2 DNA (iBB4+iBB11+iBB9, iBB4+iBB12+iBB9)</li>
+			<li>0,5 µl enzyme 2</li>
-			<li>2 µl 10x T4 DNA ligase buffer</li>
+			<li>2,2 µl Cut Smart buffer</li>
-			<li>2 µl T4 DNA ligase</li>
+			<li>20 µl bidest. H2O</li>
-			<li>4 µl ddH2O</li>
 		</ul>
-		<p>The samples were incubated for 2h at 16 °C.</p>
+		<p>Samples:</p>
+		<p>Digestion of 1,5 µl pSB1A3 iBB48647631 with <i>Spe</i>I-HF and <i>Pst</i>I-HF</p>
+		<p>Digestion of 5 µl pSB1C3 iBB5 and iBB9 with <i>Xba</i>I and <i>Pst</i>I-HF</p>
+		<p>Digestion of 4 µl pSB1C3 iBB3 and iBB4 with <i>EcoR</i>I-HF and <i>Spe</i>I-HF.</p>
+		<p>The samples were incubated for 1 h at 37 °C.</p>
+		<p>They were purified via QIAgen Gel Extraction Kit (Qiagen). Elution into 20 µl bidest. H2O.</p>
 	</div>
 </fieldset>
-<fieldset class="experiment transformation">
+<!-- Ligation -->
-     <legend>Transformation</legend>
+<fieldset class="experiment ligation">
+     <legend><a name="lig">Ligation</a></legend>
      <div class="investigator">
 		<span class="inv">Investigator:</span>
-		<span class="inv-names">Dominik</span>
+		<span class="inv-names">Christian</span>
 	</div>
      <div class="aim">
 		<span class="aim">Aim:</span>
-		<span class="aim-desc">Transformation of <i>E. coli</i> DH5&#945; with the plasmid DNA pSB1A3-iBB4+iBB11+iBB96315 and pSB1A3-iBB4+iBB12+iBB96315.</span>
+		<span class="aim-desc">Construction of P<sub>NR</sub>-test-vector, P<sub>fcpB</sub>-test-vector and antibody-vector &rarr; Ligation of pSB1A3 iBB48647631 and single BioBricks in pSB1A3.</span>
 	</div>
 	<div class="exp-content">
-		<p>The chemo competent <i>E. coli</i> DH5&alpha; cells were transformed with the plasmids pSB1A3-iBB4+iBB11+iBB96315 and pSB1A3-iBB4+iBB12+iBB96315 DNA and plated on LB-Amp-plates.<br />
+		<p>Ligation of pSB1A3 iBB48647631 with iBB5, pSB1A3 iBB48647631 with iBB3 and iBB9, pSB1A3 iBB48647631 with iBB4 and iBB9; overnight at 16°C</p>
-		The plates were incubated over night at 37° C.</p>
 	</div>
 </fieldset>
 </div>
 <div class="notebooky-entry">
 <h2 class="title">
-	<a name="25-07-2013">25.07.2013</a>
+	<a name="11-06-2013">11.06.2013</a>
 </h2>
-<fieldset class="experiment sequencing">
+<!-- Transformation-->
-     <legend>Sequencing</legend>
+<fieldset class="experiment transformation">
+     <legend>Transformation</legend>
      <div class="investigator">
 		<span class="inv">Investigator:</span>
-		<span class="inv-names">Dominik</span>
+		<span class="inv-names">Franzi</span>
 	</div>
      <div class="aim">
 		<span class="aim">Aim:</span>
-		<span class="aim-desc">Complete sequencing of pSB1A3-iBB496315.</span>
+		<span class="aim-desc">Construction of P<sub>NR</sub>-test-vector, P<sub>fcpB</sub>-test-vector and antibody-vector %rarr; Transformation of ligations.</span>
 	</div>
 	<div class="exp-content">
-		<p>8 new sequencing samples were sent out.</p>
+		<p>Transformation of the ligations into <i>E. coli</i> DH5&alpha; (30 min on ice, 60 sec 42 °C, 25 min 37 °C + 900 µl LB), plated on LB<sub>amp</sub>, overnight at 37 °C.</p>
 	</div>
 </fieldset>
-<fieldset class="experiment miniprep">
+</div>
-    <legend><a name="min">Miniprep</a></legend>
-    <div class="investigator">
-		<span class="inv">Investigator:</span>
-		<span class="inv-names">Dominik</span>
-	</div>
-    <div class="aim">
-		<span class="aim">Aim:</span>
-		<span class="aim-desc">preparation of the plasmids pSB1A3-iBB4+iBB10+iBB9 and pSB1A3-iBB4+iBB13+iBB9.</span>
-	</div>
-	<div class="exp-content">
-		<p>The plasmids were isolated with the “QIAprep Spin Miniprep Kit” (Qiagen, Düsseldorf) according to the producer instructions.</p>
-	</div>
-</fieldset>
-<fieldset class="experiment digest">
-    <legend><a name="dig">Digest</a></legend>
-    <div class="investigator">
-		<span class="inv">Investigator:</span>
-		<span class="inv-names">Dominik</span>
-	</div>
-    <div class="aim">
-		<span class="aim">Aim:</span>
-		<span class="aim-desc">Test digest of pSB1C3-iBB4+iBB10+iBB9 and pSB1C3-iBB4+iBB13+iBB9 with <i>Eco</i>RI and <i>Pst</i>I</span>
-	</div>
-	<div class="exp-content">
-		<p><ul class="digest">
-			<li>1 µl Plasmid (pSB1C3-iBB4+iBB10+iBB9 and pSB1C3-iBB4+iBB13+iBB9)</li>
-			<li>0.3 µl EcoRI</li>
-			<li>0.3 µl PstI</li>
-			<li>1 µl CutSmart</li>
-			<li>7.4 µl ddH2O</li>
-		</ul>
-		<p>The samples were incubated for 1 h at 37° C.</p>
-		<table class="gel digest">
-			<colgroup>
-				<col width="50%" />
-				<col width="50%" />
-			</colgroup>
-		<thead>
-			<tr>
-				<th colspan="2" class="title">Gel electrophoresis</th>
-			</tr>
-		</thead>
-		<tbody>
-			<tr>
-				<td>
-					<img src="beispiel.png" width="50%" alt="gel-electrophoresis-image" />
-				</td>
-				<td>
-					<p>
-					<span class="gel-elc">Gel substances</span>
-					<ul class="gel-sub">
-						<li>1% Agarose gel</li>
-						<li>10 µl RedSafe in 50 ml gel</li>
-						<li>x µl Hyper Ladder</li>
-					</ul>
-					</p>
-					<p>
-						<span class="exp">Expactations</span>
-						<ul class="exp">
-							<li>Lane 1: 5300 kbp</li>
-							<li>Lane 2: 3300 kbp</li>
-							<li>Lane 3: 1300 kbp</li>
-						</ul>
-					</p>
-					<p>Two samples of each plasmid preparation showed the expected fragments.</p>
-				</td>
-			</tr>
-		</tbody>
-		</table>
-	</div>
-</fieldset>
+<div class="notebooky-entry">
+<h2 class="title">
+	<a name="12-06-2013">12.06.2013</a>
+</h2>
+<!-- Inoculation -->
 <fieldset class="experiment inoculation">
      <legend><a name="ino">Inoculation</a></legend>
      <div class="investigator">
 		<span class="inv">Investigator:</span>
-		<span class="inv-names">Dominik</span>
+		<span class="inv-names">Franzi</span>
 	</div>
      <div class="aim">
 		<span class="aim">Aim:</span>
-		<span class="aim-desc">inoculation of colonies for plasmid preparation</span>
+		<span class="aim-desc">Construction of P<sub>NR</sub>-test-vector, P<sub>fcpB</sub>-test-vector and antibody-vector &rarr; Inoculation of transformants for miniprep.</span>
 	</div>
 	<div class="exp-content">
-		<p>4 colonies of pSB1A3-iBB4+iBB11+iBB96315 and 4 colonies of pSB1A3-iBB4+iBB12+iBB96315 were inoculated in 5 ml LB medium containing chloramphenicol.</p>
+		<p>4 colonies of each transformation inoculated in 4 ml LB<sub>amp</sub>, overnight at 37 °C.</p>
 	</div>
 </fieldset>
-<fieldset class="experiment competent cells">
+</div>
-     <legend>Competent cells</legend>
+<div class="notebooky-entry">
+<h2 class="title">
+	<a name="13-06-2013">13.06.2013</a>
+</h2>
+<!-- Miniprep -->
+<fieldset class="experiment miniprep">
+     <legend><a name="min">Miniprep</a></legend>
      <div class="investigator">
 		<span class="inv">Investigator:</span>
-		<span class="inv-names">Dominik</span>
+		<span class="inv-names">Franzi</span>
 	</div>
      <div class="aim">
 		<span class="aim">Aim:</span>
-		<span class="aim-desc">Overnight culture for making new aliquots of competent cells.</span>
+		<span class="aim-desc">Construction of P<sub>NR</sub>-test-vector, P<sub>fcpB</sub>-test-vector and antibody-vector &rarr; Miniprep of transformants.</span>
 	</div>
 	<div class="exp-content">
-		<p>For making new aliquots of <i>E. coli</i> DH5α cells 50 ml LB medium were inoculated with 500 µl of an <i>E. coli</i> DH5α culture.</p>
+		<p>Miniprep of the liquid cultures (“QIAprep Spin Miniprep Kit” (Qiagen, Düsseldorf)), eluted in 30 µl bidest. H2O.</p>
 	</div>
 </fieldset>
-</div>
-<div class="notebooky-entry">
-<h2 class="title">
-	<a name="26-07-2013">26.07.2013</a>
-</h2>
+<!-- Digest -->
 <fieldset class="experiment digest">
      <legend><a name="dig">Digest</a></legend>
      <div class="investigator">
 		<span class="inv">Investigator:</span>
-		<span class="inv-names">Dominik</span>
+		<span class="inv-names">Franzi</span>
 	</div>
      <div class="aim">
 		<span class="aim">Aim:</span>
-		<span class="aim-desc">Digest of pSB1C3-iBB4+iBB10+iBB9, pSB1C3-iBB4+iBB13+iBB9, pSB1C3-iBB6315 and pSB1A3.</span>
+		<span class="aim-desc">Construction of P<sub>NR</sub>-test-vector, P<sub>fcpB</sub>-test-vector and antibody-vector &rarr; Control digestion the minipreps and pSB1A3 iBB48647631 K3 (comparison of size).</span>
 	</div>
 	<div class="exp-content">
-		<p><ul class="digest">
+		<ul class="digest">
-			<li>1000 ng Plasmid (pSB1C3-iBB4+iBB10+iBB9, pSB1C3-iBB4+iBB13+iBB9)</li>
+			<li>1 µl DNA template</li>
-			<li>0.5 µl EcoRI</li>
+			<li>0,3 µl <i>EcoR</i>I-HF</li>
-			<li>0.5 µl SpeI</li>
+			<li>0,3 µl <i>Pst</i>I-HF</li>
-			<li>2 µl CutSmart</li>
+			<li>1 µl Cut Smart buffer</li>
-			<li>ad 20 µl ddH2O</li>
+			<li>7,4 µl bidest. H2O</li>
 		</ul>
+		<p>The samples were incubated for 1 h at 37 °C.</p>
-		<p><ul class="digest">
-			<li>1000 ng Plasmid (pSB1C3-iBB6315)</li>
-			<li>0.5 µl XbaI</li>
-			<li>0.5 µl PstI</li>
-			<li>2 µl CutSmart</li>
-			<li>ad 20 µl ddH2O</li>
-		</ul>
-		<p><ul class="digest">
-			<li>1000 ng Plasmid (pSB1A3)</li>
-			<li>0.5 µl EcoRI</li>
-			<li>0.5 µl PstI</li>
-			<li>2 µl CutSmart</li>
-			<li>ad 20 µl ddH2O</li>
-		</ul>
-		<p>The samples were incubated for 1 h at 37° C.</p>
 		<table class="gel digest">
 			<colgroup>
@@ Line 998: / Line 895: @@
 			<tr>
 				<td>
-					<img src="beispiel.png" width="50%" alt="gel-electrophoresis-image" />
+					<img src="https://static.igem.org/mediawiki/2013/2/22/Gel_2013-06-13_1.png" width="100%" alt="gel-electrophoresis-image" />
 				</td>
 				<td>
@@ Line 1,005: / Line 902: @@
 					<ul class="gel-sub">
 						<li>1% Agarose gel</li>
-						<li>10 µl RedSafe in 50 ml gel</li>
+						<li>10 µl RedSafe per 50 ml gel</li>
-						<li>x µl Hyper Ladder</li>
+						<li>6 µl 2-Log DNA Ladder (NEB)</li>
+						<li>6x loading buffer (for samples)</li>
 					</ul>
 					</p>
 					<p>
-						<span class="exp">Expactations</span>
+							<table class="gel">
-						<ul class="exp">
+								<colgroup>
-							<li>Lane 1: 5300 kbp</li>
+									<col width="10%" />
-							<li>Lane 2: 3300 kbp</li>
+									<col width="2%" />
-							<li>Lane 3: 1300 kbp</li>
+									<col width="50%" />
-						</ul>
+									<col width="5%" />
+									<col width="33%" />
+								</colgroup>
+								<thead>
+									<th>Lane</th>
+									<th>&nbsp;</th>
+									<th>Content</th>
+									<th>&nbsp;</th>
+									<th>Expectations</th>
+								</thead>
+								<tr>
+									<td>upper:</td>
+								</tr>
+								<tr>
+									<td>2+3</td>
+									<th>&nbsp;</th>
+									<td>iBB49 K3-4</td>
+									<th>&nbsp;</th>
+									<td>1260 bp</td>
+								</tr>
+								<tr>
+									<td>4-7</td>
+									<th>&nbsp;</th>
+									<td>iBB39 K1-4</td>
+									<th>&nbsp;</th>
+									<td>1030 bp</td>
+								</tr>
+								<tr>
+									<td>lower:</td>
+								</tr>
+								<tr>
+									<td>2</td>
+									<th>&nbsp;</th>
+									<td>iBB49647631</td>
+									<th>&nbsp;</th>
+									<td>4157 bp</td>
+								</tr>
+								<tr>
+									<td>3-6</td>
+									<th>&nbsp;</th>
+									<td>iBB496476315 K1-4</td>
+									<th>&nbsp;</th>
+									<td>4404 bp</td>
+								</tr>
+								<tr>
+									<td>7+8</td>
+									<th>&nbsp;</th>
+									<td>iBB49 K1-2</td>
+									<th>&nbsp;</th>
+									<td>1260 bp</td>
+								</tr>
+								<tr>
+									<td></td>
+									<th>&nbsp;</th>
+									<td></td>
+									<th>&nbsp;</th>
+									<td>Vector pSB1A3: 2150 bp</td>
+								</tr>
+							</table>
 					</p>
-					<p>All expected fragments were present. The relevant fragments were cut from the gel and then purified via “Qiagen Gel Extraction Kit” (Qiagen, Düsseldorf).</p>
+					<p>IBB48647631 and iBB486476315 probably positive, but better repeat or do a Colony-PCR.</p>
 				</td>
 			</tr>
@@ Line 1,025: / Line 981: @@
 </fieldset>
-<fieldset class="experiment ligation">
+</div>
-    <legend><a name="lig">Ligation</a></legend>
-    <div class="investigator">
-		<span class="inv">Investigator:</span>
-		<span class="inv-names">Dominik</span>
-	</div>
-    <div class="aim">
-		<span class="aim">Aim:</span>
-		<span class="aim-desc">Assemble the biobricks iBB4+iBB10+iBB9, iBB4+iBB13+iBB9 and iBB96315 into the vector pSB1A3.</span>
-	</div>
-	<div class="exp-content">
-		<p><ul class="lig">
-			<li>2 µl vector DNA (pSB1A3)</li>
-			<li>5 µl insert 1 DNA (iBB96315)</li>
-			<li>5 µl insert 2 DNA (iBB4+iBB10+iBB9,iBB4+iBB13+iBB9)</li>
-			<li>2 µl 10x T4 DNA ligase buffer</li>
-			<li>2 µl T4 DNA ligase</li>
-			<li>4 µl ddH2O</li>
-		</ul>
-		<p>The samples were incubated for 1h at 16 °C.</p>
-		<p>Resulting in the plasmids pSB1A3-iBB4+iBB10+iBB96315 and pSB1A3-iBB4+iBB13+iBB96315.</p>
-	</div>
-</fieldset>
-<fieldset class="experiment miniprep">
-    <legend><a name="min">Miniprep</a></legend>
-    <div class="investigator">
-		<span class="inv">Investigator:</span>
-		<span class="inv-names">Dominik</span>
-	</div>
-    <div class="aim">
-		<span class="aim">Aim:</span>
-		<span class="aim-desc">preparation of the pSB1A3-iBB4+iBB11+iBB96315 and pSB1A3-iBB4+iBB12+iBB96315.</span>
-	</div>
-	<div class="exp-content">
-		<p>The plasmids were isolated with the “QIAprep Spin Miniprep Kit” (Qiagen, Düsseldorf) according to the producer instructions.</p>
-	</div>
-</fieldset>
+<div class="notebooky-entry">
+<h2 class="title">
+	<a name="16-06-2013">16.06.2013</a>
+</h2>
+<!-- Digest -->
 <fieldset class="experiment digest">
      <legend><a name="dig">Digest</a></legend>
      <div class="investigator">
 		<span class="inv">Investigator:</span>
-		<span class="inv-names">Dominik</span>
+		<span class="inv-names">Lukas</span>
 	</div>
      <div class="aim">
 		<span class="aim">Aim:</span>
-		<span class="aim-desc">Test digest of ppSB1A3-iBB4+iBB11+iBB96315 and pSB1A3-iBB4+iBB12+iBB96315 with <i>Eco</i>RI and <i>Pst</i>I</span>
+		<span class="aim-desc">Construction of antibody-vector &rarr; Repeat of control digestion.</span>
 	</div>
 	<div class="exp-content">
-		<p><ul class="digest">
+		<ul class="digest">
-			<li>1 µl Plasmid (pSB1C3-iBB39,pSB1A3-iBB49 and pSB1C3-iBB6315)</li>
+			<li>1 µl DNA template</li>
-			<li>0.3 µl EcoRI</li>
+			<li>0,3 µl <i>EcoR</i>I-HF</li>
-			<li>0.3 µl PstI</li>
+			<li>0,3 µl <i>Pst</i>I-HF</li>
-			<li>1 µl CutSmart</li>
+			<li>1 µl Cut Smart buffer</li>
-			<li>7.4 µl ddH2O</li>
+			<li>7,4 µl bidest. H2O</li>
 		</ul>
-		<p>The samples were incubated for 1 h at 37° C.</p>
+		<p>Templates: PSB1A3 iBB486476315 and pSB1A3 iBB48647631 K3 (comparison of size).</p>
+		<p>The samples were incubated for 1 h at 37 °C.</p>
 		<table class="gel digest">
 			<colgroup>
@@ Line 1,096: / Line 1,023: @@
 			<tr>
 				<td>
-					<img src="beispiel.png" width="50%" alt="gel-electrophoresis-image" />
+					<img src="https://static.igem.org/mediawiki/2013/3/38/Gel_2013-06-16.png" width="100%" alt="gel-electrophoresis-image" />
 				</td>
 				<td>
@@ Line 1,103: / Line 1,030: @@
 					<ul class="gel-sub">
 						<li>1% Agarose gel</li>
-						<li>10 µl RedSafe in 50 ml gel</li>
+						<li>10 µl RedSafe per 50 ml gel</li>
-						<li>x µl Hyper Ladder</li>
+						<li>6 µl GeneRuler&trade; 1kb plus DNA Ladder (Thermo Scientific)</li>
+						<li>6x loading buffer (for samples)</li>
 					</ul>
 					</p>
 					<p>
-						<span class="exp">Expactations</span>
+							<table class="gel">
-						<ul class="exp">
+								<colgroup>
-							<li>Lane 1: 5300 kbp</li>
+									<col width="10%" />
-							<li>Lane 2: 3300 kbp</li>
+									<col width="2%" />
-							<li>Lane 3: 1300 kbp</li>
+									<col width="50%" />
-						</ul>
+									<col width="5%" />
+									<col width="33%" />
+								</colgroup>
+								<thead>
+									<th>Lane</th>
+									<th>&nbsp;</th>
+									<th>Content</th>
+									<th>&nbsp;</th>
+									<th>Expectations</th>
+								</thead>
+								<tr>
+									<td>2+3</td>
+									<th>&nbsp;</th>
+									<td>iBB48647631</td>
+									<th>&nbsp;</th>
+									<td>4157 bp</td>
+								</tr>
+								<tr>
+									<td>4-7</td>
+									<th>&nbsp;</th>
+									<td>iBB486476315</td>
+									<th>&nbsp;</th>
+									<td>TBA bp</td>
+								</tr>
+								<tr>
+									<td></td>
+									<th>&nbsp;</th>
+									<td></td>
+									<th>&nbsp;</th>
+									<td>Vector pSB1A3: 2150 bp</td>
+								</tr>
+							</table>
 					</p>
-					<p>Most of the preparations resulted in the expected fragments, all others were discarded.</p>
+					<p>Too much DNA used, will be repeated as PCR.</p>
 				</td>
 			</tr>
@@ Line 1,123: / Line 1,082: @@
 </fieldset>
-<fieldset class="experiment competent cells">
-    <legend>Competent cells</legend>
-    <div class="investigator">
-		<span class="inv">Investigator:</span>
-		<span class="inv-names">Dominik</span>
-	</div>
-    <div class="aim">
-		<span class="aim">Aim:</span>
-		<span class="aim-desc">New aliquots of competent cells (<i>E. coli</i> DH5α).</span>
-	</div>
-	<div class="exp-content">
-		<p>About 80 new aliquots were made.</p>
-	</div>
-</fieldset>
 </div>
 <div class="notebooky-entry">
 <h2 class="title">
-	<a name="02-07-2013">02.07.2013</a>
+	<a name="17-06-2013">17.06.2013</a>
 </h2>
-<fieldset class="experiment sequencing">
+<fieldset class="experiment digest">
-     <legend>Sequencing</legend>
+     <legend><a name="dig">Digest</a></legend>
      <div class="investigator">
 		<span class="inv">Investigator:</span>
-		<span class="inv-names">Dominik</span>
+		<span class="inv-names">Alex</span>
 	</div>
      <div class="aim">
 		<span class="aim">Aim:</span>
-		<span class="aim-desc">Analysis of the sequence of pSB1A3-iBB496315.</span>
+		<span class="aim-desc">Digest of pSB1A3_3+9 and pSB1A3_4+9 with <i>Spe</i>I and <i>Pst</i>I, and corresponding inserts pSB1C3_5+4+1+6 and pSB1C3_6+3+1+5 with <i>EcoR</i>I and <i>Pst</i>I.</span>
 	</div>
 	<div class="exp-content">
-		<p>Obviously the wrong plasmid was sent out for sequencing. Due to the lack of enough plasmids for an additional sequence analysis, we will redo a new miniprep.</p>
+		<ul class="digest">
-	</div>
+ 			<li>1 µl DNA</li>
+			<li>0.3 µl Enzyme 1 (<i>Spe</i>I or <i>EcoR</i>I, resp.)</li>
+			<li>0.3 µl Enzyme 2 (<i>Pst</i>I)</li>
+			<li>1 µl Cut Smart buffer</li>
+			<li>7.4 µl ddbidest. H2O</li>
+		</ul>
+		<p>The samples were incubated for 1.5 h at 37 °C.</p>
+		<p>Approaches were unsewed by means of gel electrophoresis, corresponding bands were picked under UV, DNA extraction from gel was performed using DNA Gel Extraction Kit (<i>QIAGEN</i>).</p>
 </fieldset>
-<fieldset class="experiment transformation">
+<fieldset class="experiment ligation">
-     <legend>Transformation</legend>
+     <legend><a name="lig">Ligation, Transformation, Plasmid prep</a></legend>
      <div class="investigator">
 		<span class="inv">Investigator:</span>
-		<span class="inv-names">Dominik</span>
+		<span class="inv-names">Alex</span>
 	</div>
      <div class="aim">
 		<span class="aim">Aim:</span>
-		<span class="aim-desc">Transformation of <i>E. coli</i> DH5&#945; with the plasmid DNA pSB1A3-iBB4+iBB10+iBB96315 and pSB1A3-iBB4+iBB13+iBB96315.</span>
+		<span class="aim-desc">Assemble the combined bricks 5416 and 6315 into their appropriate vectors pSB1A3_39 and pSB1A3_49, resp.</span>
 	</div>
 	<div class="exp-content">
-		<p>The chemo competent <i>E. coli</i> DH5&alpha; cells were transformed with the plasmids pSB1A3-iBB4+iBB10+iBB96315 and pSB1A3-iBB4+iBB13+iBB96315 and plated on LB-Amp-plates.<br />
+		<ul class="lig">
-		The plates were incubated over night at 37° C.</p>
+			<li>Insert und plasmid were transformed using a 1:2 ratio. Samples were incubated ON at RT.</li>
+			<li>Ligation product were transformed into chemical competent DH5α cells (30 min on ice, 90 sec 42 °C, 45 min 37 °C + 900 µl LB).</li>
+			<li>Transformed cells were spread out on LB Amp culture plates.</li>
+			<li>Incubation ON at RT.</li>
+                        <li>4 clones were picked for overnight at cultures.</li>
+                        <li>A plasmid preparation was carried out the next day.</li>
+		</ul>
 	</div>
 </fieldset>
@@ Line 1,178: / Line 1,137: @@
 <div class="notebooky-entry">
 <h2 class="title">
-	<a name="30-07-2013">30.07.2013</a>
+	<a name="22-06-2013">22.06.2013</a>
 </h2>
 <fieldset class="experiment digest">
-     <legend><a name="dig">Digest</a></legend>
+     <legend><a name="dig">Test digest</a></legend>
      <div class="investigator">
 		<span class="inv">Investigator:</span>
-		<span class="inv-names">Dominik</span>
+		<span class="inv-names">Alex</span>
 	</div>
      <div class="aim">
 		<span class="aim">Aim:</span>
-		<span class="aim-desc">Digest of pSB1A3-iBB49 and pSB1C3-iBB6315.</span>
+		<span class="aim-desc">Digest of pSB1A3_395416 and pSB1A3_496315 with <i>EcoR</i>I and <i>Pst</i>I.</span>
 	</div>
 	<div class="exp-content">
-		<p><ul class="digest">
+		<ul class="digest">
-			<li>1000 ng Plasmid (pSB1A3-iBB49)</li>
+			<li>1 µl Plasmid DNA</li>
-			<li>0.5 µl PstI</li>
+			<li>0.3 µl EcoRI</li>
-			<li>0.5 µl SpeI</li>
+			<li>0.3 µL PstI</li>
-			<li>2 µl CutSmart</li>
+			<li>1 µl Cut Smart</li>
-			<li>ad 20 µl ddH2O</li>
+			<li>7.4 µl ddbidest. H2O</li>
 		</ul>
+		<p>The samples were incubated for 2 h at 37 °C.</p>
-		<p><ul class="digest">
-			<li>1000 ng Plasmid (pSB1C3-iBB6315)</li>
-			<li>0.5 µl XbaI</li>
-			<li>0.5 µl PstI</li>
-			<li>2 µl CutSmart</li>
-			<li>ad 20 µl ddH2O</li>
-		</ul>
-		<p>The samples were incubated for 1 h at 37° C.</p>
 		<table class="gel digest">
 			<colgroup>
@@ Line 1,222: / Line 1,172: @@
 			<tr>
 				<td>
-					<img src="beispiel.png" width="50%" alt="gel-electrophoresis-image" />
+					<img src="https://static.igem.org/mediawiki/2013/5/53/Gel_2013-06-17_2.png" width="100%" alt="gel-electrophoresis-image" />
 				</td>
 				<td>
@@ Line 1,229: / Line 1,179: @@
 					<ul class="gel-sub">
 						<li>1% Agarose gel</li>
-						<li>10 µl RedSafe in 50 ml gel</li>
+						<li>10 µl Ethidium bromide 50 ml gel</li>
-						<li>x µl Hyper Ladder</li>
+						<li>4 µl 2-log DNA ladder (New England Biolabs)</li>
 					</ul>
 					</p>
 					<p>
-						<span class="exp">Expactations</span>
+							<table class="gel">
-						<ul class="exp">
+								<colgroup>
-							<li>Lane 1: 5300 kbp</li>
+									<col width="10%" />
-							<li>Lane 2: 3300 kbp</li>
+									<col width="2%" />
-							<li>Lane 3: 1300 kbp</li>
+									<col width="50%" />
-						</ul>
+									<col width="5%" />
+									<col width="33%" />
+								</colgroup>
+								<thead>
+									<th>Lane</th>
+									<th>&nbsp;</th>
+									<th>Content</th>
+									<th>&nbsp;</th>
+									<th>Expectations</th>
+								</thead>
+								<tr>
+									<td>upper:</td>
+								</tr>
+								<tr>
+									<td>2</td>
+									<th>&nbsp;</th>
+									<td>iBB6315</td>
+									<th>&nbsp;</th>
+									<td>1200 + 2000 bp</td>
+								</tr>
+								<tr>
+									<td>3</td>
+									<th>&nbsp;</th>
+									<td>iBB49</td>
+									<th>&nbsp;</th>
+									<td>1200 + 2000 bp</td>
+								</tr>
+								<tr>
+								<tr>
+									<td>4-7</td>
+									<th>&nbsp;</th>
+									<td>iBB496315 K4-1</td>
+									<th>&nbsp;</th>
+									<td>2400 + 2000 bp</td>
+								</tr>
+								<tr>
+									<td>lower:</td>
+								</tr>
+								<tr>
+									<td>2</td>
+									<th>&nbsp;</th>
+									<td>iBB5416</td>
+									<th>&nbsp;</th>
+									<td>1500 + 2000 bp</td>
+								</tr>
+								<tr>
+									<td>3</td>
+									<th>&nbsp;</th>
+									<td>iBB39</td>
+									<th>&nbsp;</th>
+									<td>1000 + 2000 bp</td>
+								</tr>
+								<tr>
+									<td>4-7</td>
+									<th>&nbsp;</th>
+									<td>iBB395416 K1-4</td>
+									<th>&nbsp;</th>
+									<td>2400 + 2000 bp</td>
+								</tr>
+							</table>
 					</p>
-					<p>All expected fragments were present. The relevant fragments were cut from the gel and then purified via “Qiagen Gel Extraction Kit” (Qiagen, Düsseldorf).</p>
+				</td>
+			</tr>
+			<tr>
+				<td colspan="2" class="gel-fazit">
+					<p>We didn't receive all expected fragments. All expected fragments in lanes 4-7 were missing. Ligation failed &#x2639;</p>
 				</td>
 			</tr>
@@ Line 1,249: / Line 1,262: @@
 </fieldset>
-<fieldset class="experiment inoculation">
-    <legend><a name="ino">Inoculation</a></legend>
-    <div class="investigator">
-		<span class="inv">Investigator:</span>
-		<span class="inv-names">Dominik</span>
-	</div>
-    <div class="aim">
-		<span class="aim">Aim:</span>
-		<span class="aim-desc">inoculation of colonies for plasmid preparation</span>
-	</div>
-	<div class="exp-content">
-		<p>4 colonies of pSB1A3-iBB4+iBB10+iBB96315 and the only single colony of pSB1A3-iBB4+iBB13+iBB96315 were inoculated in 5 ml LB medium containing chloramphenicol. <font color="#FF0000">Additionally the remaining ligation preparation was transformed in <i>E. coli</i> DH5α?????.</font></p>
-	</div>
-</fieldset>
 </div>
 <div class="notebooky-entry">
 <h2 class="title">
-	<a name="31-07-2013">31.07.2013</a>
+	<a name="24-06-2013">24.06.2013</a>
 </h2>
-<fieldset class="experiment ligation">
+<fieldset class="experiment pcr">
-     <legend><a name="lig">Ligation</a></legend>
+     <legend><a name="pcr">PCR</a></legend>
      <div class="investigator">
 		<span class="inv">Investigator:</span>
-		<span class="inv-names">Dominik</span>
+		<span class="inv-names">Alex</span>
 	</div>
      <div class="aim">
 		<span class="aim">Aim:</span>
-		<span class="aim-desc">Ligation of pSB1A3-iBB49 and iBB6315.</span>
+		<span class="aim-desc">Check right assembly of BioBrick BBa_K1071005 in the plasmid for antibody production</span>
 	</div>
 	<div class="exp-content">
-		<p><ul class="lig">
+		<table class="pcr">
-			<li>2 µl vector DNA (pSB1A3-iBB49) (40 ng)</li>
+			<colgroup>
-			<li>5 µl insert DNA (iBB6315) (150 ng)</li>
+				<col width="15%" />
-			<li>2 µl 10x T4 DNA ligase buffer</li>
+				<col width="30%" />
-			<li>2 µl T4 DNA ligase</li>
+				<col width="5%" />
-			<li>9 µl ddH2O</li>
+				<col width="20%" />
-		</ul>
+				<col width="20%" />
-		<p>The samples were incubated for 1h at 16 °C.</p>
+				<col width="10%" />
-	</div>
+			</colgroup>
-</fieldset>
+			<thead>
+				<th>Volume</th>
-<fieldset class="experiment transformation">
+				<th>Reagent</th>
-    <legend>Transformation</legend>
+				<th>&nbsp;</th>
-    <div class="investigator">
+				<th>Temp (°C)</th>
-		<span class="inv">Investigator:</span>
+				<th colspan="2">Time</th>
-		<span class="inv-names">Dominik</span>
+			</thead>
-	</div>
+			<tr>
-    <div class="aim">
+				<td>1 µl</td>
-		<span class="aim">Aim:</span>
+				<td>pSB1A3_486476315</td>
-		<span class="aim-desc">Transformation of <i>E. coli</i> DH5&#945; with the plasmid DNA pSB1A3-iBB49+iBB6315.</span>
+				<td>&nbsp;</td>
-	</div>
+				<td>95</td>
-	<div class="exp-content">
+				<td colspan="2">3 min</td>
-		<p>The chemo competent <i>E. coli</i> DH5&alpha; cells were transformed with the plasmid pSB1A3-iBB49+iBB6315 and plated on LB-Amp-plates.<br />
+			</tr>
-		The plates were incubated over night at 37° C.</p>
+			<tr>
-	</div>
+				<td>0.5 µl</td>
-</fieldset>
+				<td>Primer fwd</td>
+				<td>&nbsp;</td>
-<fieldset class="experiment miniprep">
+				<td>95</td>
-    <legend><a name="min">Miniprep</a></legend>
+				<td colspan="2">30 sec</td>
-    <div class="investigator">
+			</tr>
-		<span class="inv">Investigator:</span>
+			<tr>
-		<span class="inv-names">Dominik</span>
+				<td>0.5 µl</td>
-	</div>
+				<td>Primer rev</td>
-    <div class="aim">
+				<td>&nbsp;</td>
-		<span class="aim">Aim:</span>
+				<td>60</td>
-		<span class="aim-desc">Preparation of the pSB1A3-iBB4+iBB10+iBB96315 and pSB1A3-iBB4+iBB13+iBB96315.</span>
+				<td>30 sec</td>
-	</div>
+				<td rowspan="3">x30</td>
-	<div class="exp-content">
+			</tr>
-		<p>The culture of pSB1A3-iBB4+iBB13+iBB96315 had a pinkish color and therefore was discarded. The plasmid pSB1A3-iBB4+iBB10+iBB96315 was isolated with the “QIAprep Spin Miniprep Kit” (Qiagen, Düsseldorf) according to the producer instructions.</p>
+			<tr>
-	</div>
+				<td>0.5 µl</td>
-</fieldset>
+				<td>dNTPs</td>
+				<td>&nbsp;</td>
-<fieldset class="experiment digest">
+				<td>72</td>
-    <legend><a name="dig">Digest</a></legend>
+				<td>42 sec</td>
-    <div class="investigator">
+			</tr>
-		<span class="inv">Investigator:</span>
+			<tr>
-		<span class="inv-names">Dominik</span>
+				<td>0.5 µl</td>
-	</div>
+				<td>Phusion polymerase</td>
-    <div class="aim">
+				<td>&nbsp;</td>
-		<span class="aim">Aim:</span>
+				<td>72</td>
-		<span class="aim-desc">Test digest of pSB1A3-iBB4+iBB10+iBB96315 with <i>Eco</i>RI and <i>Pst</i>I</span>
+				<td>5 min</td>
-	</div>
+			</tr>
-	<div class="exp-content">
+			<tr>
-		<p><ul class="digest">
+				<td>5 µl</td>
-			<li>1 µl Plasmid (pSB1A3-iBB4+iBB10+iBB96315)</li>
+				<td>Phusion buffer 5x</td>
-			<li>0.3 µl EcoRI</li>
+				<td>&nbsp;</td>
-			<li>0.3 µl PstI</li>
+				<td>4</td>
-			<li>1 µl CutSmart</li>
+				<td colspan="2"><span class="hold">Hold</span></td>
-			<li>7.4 µl ddH2O</li>
+			</tr>
-		</ul>
+                        <tr>
-		<p>The samples were incubated for 1 h at 37° C.</p>
+				<td>add 25 µL</td>
-		<table class="gel digest">
+				<td>ddbidest. H2O</td>
+				<td>&nbsp;</td>
+			</tr>
+		</table>
+		<br />
+		<!-- Das Gel-Bild fast gleich wie beim Verdau -->
+		<table class="gel pcr">
 			<colgroup>
 				<col width="50%" />
@@ Line 1,355: / Line 1,360: @@
 			<tr>
 				<td>
-					<img src="beispiel.png" width="50%" alt="gel-electrophoresis-image" />
+					<img src="https://static.igem.org/mediawiki/2013/5/58/Gel_2013-06-24.png" width="100%" alt="gel-electrophoresis-image" />
 				</td>
 				<td>
@@ Line 1,362: / Line 1,367: @@
 					<ul class="gel-sub">
 						<li>1% Agarose gel</li>
-						<li>10 µl RedSafe in 50 ml gel</li>
+						<li>10 µl Ethidium bromide 50 ml gel</li>
-						<li>x µl Hyper Ladder</li>
+						<li>4 µl 2-log DNA ladder (New England Biolabs)</li>
 					</ul>
 					</p>
 					<p>
-						<span class="exp">Expactations</span>
+							<table class="gel">
-						<ul class="exp">
+								<colgroup>
-							<li>Lane 1: 5300 kbp</li>
+									<col width="10%" />
-							<li>Lane 2: 3300 kbp</li>
+									<col width="2%" />
-							<li>Lane 3: 1300 kbp</li>
+									<col width="50%" />
-						</ul>
+									<col width="5%" />
+									<col width="33%" />
+								</colgroup>
+								<thead>
+									<th>Lane</th>
+									<th>&nbsp;</th>
+									<th>Content</th>
+									<th>&nbsp;</th>
+									<th>Expectations</th>
+								</thead>
+								<tr>
+									<td>2-5</td>
+									<th>&nbsp;</th>
+									<td>pSB1A3 iBB486476315 K1-4</td>
+									<th>&nbsp;</th>
+									<td>
+										<p>~ 6,000 bp for pSB1A3_486476315</p>
+										<p>280 bp for BBa_K1071005</p>
+									</td>
+								</tr>
+							</table>
 					</p>
-					<p>Worked as expected.</p>
+					<p>Each sample contained 2 bands, that means we got all expected bands and it could be assumed that the insertion of BBa_K1071005 was successful.</p>
 				</td>
 			</tr>
@@ Line 1,382: / Line 1,407: @@
 </fieldset>
-<fieldset class="experiment sequencing">
+</div>
-    <legend>Sequencing</legend>
-    <div class="investigator">
-		<span class="inv">Investigator:</span>
-		<span class="inv-names">Dominik</span>
-	</div>
-    <div class="aim">
-		<span class="aim">Aim:</span>
-		<span class="aim-desc">Examination of the sequence of pSB1A3-iBB4+iBB10+iBB96315.</span>
-	</div>
-	<div class="exp-content">
-		<p>8 samples were sent out for sequencing.</p>
-	</div>
-</fieldset>
-<fieldset class="experiment inoculation">
+<div class="notebooky-entry">
-     <legend><a name="ino">Inoculation</a></legend>
+<h2 class="title">
+	<a name="25-06-2013">25.06.2013</a>
+</h2>
+<fieldset class="experiment digest">
+     <legend><a name="dig">Digest</a></legend>
      <div class="investigator">
 		<span class="inv">Investigator:</span>
@@ Line 1,405: / Line 1,422: @@
      <div class="aim">
 		<span class="aim">Aim:</span>
-		<span class="aim-desc">inoculation of colonies for plasmid preparation</span>
+		<span class="aim-desc">Test digest of pSB1A3-iBB39, pSB1A3-iBB49 and pSB1C3-iBB6315 with <i>EcoR</i>I and <i>Pst</i>I</span>
 	</div>
 	<div class="exp-content">
-		<p>Both colonies of pSB1A3-iBB4+iBB13+iBB96315 were inoculated in 5 ml LB medium containing ampicillin.</p>
+		<p><ul class="digest">
+			<li>1 µl Plasmid (pSB1C3-iBB39,pSB1A3-iBB49 and pSB1C3-iBB6315)</li>
+			<li>0.3 µl <i>EcoR</i>I</li>
+			<li>0.3 µl <i>Pst</i>I</li>
+			<li>1 µl CutSmart</li>
+			<li>7.4 µl ddbidest. H2O</li>
+		</ul>
+		<p>The samples were incubated for 1 h at 37 °C.</p>
+		<p>Worked as expected.</p>
 	</div>
 </fieldset>
@@ Line 1,415: / Line 1,440: @@
 <div class="notebooky-entry">
 <h2 class="title">
-	<a name="01-08-2013">01.08.2013</a>
+	<a name="26-06-2013">26.06.2013</a>
 </h2>
-<fieldset class="experiment sequencing">
+<fieldset class="experiment digest">
-     <legend>Sequencing</legend>
+     <legend><a name="dig">Digest</a></legend>
      <div class="investigator">
 		<span class="inv">Investigator:</span>
@@ Line 1,426: / Line 1,451: @@
      <div class="aim">
 		<span class="aim">Aim:</span>
-		<span class="aim-desc">Examination of the sequence of pSB1A3-iBB4+iBB10+iBB96315.</span>
+		<span class="aim-desc">Construction of the plasmids pSB1A3-iBB10+9, pSB1A3-iBB11+9, pSB1A3-iBB12+9 and pSB1A3-iBB13+9</span>
 	</div>
 	<div class="exp-content">
-		<p>2 additional sequence samples were sent out for sequencing.</p>
+		<p><ul class="digest">
+			<li>1 µl Plasmid (pSB1C3-iBB10, pSB1C3-iBB11, pSB1C3-iBB12 and pSB1C3-iBB13)</li>
+			<li>0.5 µl <i>EcoR</i>I</li>
+			<li>0.5 µl <i>Spe</i>I</li>
+			<li>2 µl CutSmart</li>
+			<li>12 µl ddbidest. H2O</li>
+		</ul>
+		<p><ul class="digest">
+			<li>1 µl Plasmid (pSB1C3-iBB9)</li>
+			<li>0.5 µl <i>Xba</i>I</li>
+			<li>0.5 µl <i>Pst/</i>I</li>
+			<li>2 µl CutSmart</li>
+			<li>12 µl ddbidest. H2O</li>
+		</ul>
+		<p><ul class="digest">
+			<li>1 µl Plasmid (pSB1A3)</li>
+			<li>0.5 µl <i>EcoR</i>I</li>
+			<li>0.5 µl <i>Pst</i>I</li>
+			<li>2 µl CutSmart</li>
+			<li>12 µl ddbidest. H2O</li>
+		</ul>
+		<p>The samples were incubated for 1.5 h at 37 °C.</p>
+		<p>All expected fragments were present. The relevant fragments were cut from the gel and then purified via QIAgen Gel Extraction Kit (Qiagen).</p>
 	</div>
 </fieldset>
-<fieldset class="experiment digest">
+<fieldset class="experiment ligation">
-     <legend><a name="dig">Digest</a></legend>
+     <legend><a name="lig">Ligation</a></legend>
      <div class="investigator">
 		<span class="inv">Investigator:</span>
@@ Line 1,441: / Line 1,491: @@
      <div class="aim">
 		<span class="aim">Aim:</span>
-		<span class="aim-desc">Test digest of pSB1A3-iBB4+iBB13+iBB96315 with <i>Eco</i>RI and <i>Pst</i>I</span>
+		<span class="aim-desc">Assemble the biobricks iBB10+iBB9, iBB11+iBB9, iBB12+iBB9 and iBB13+iBB9 into the vector pSB1A3.</span>
 	</div>
 	<div class="exp-content">
-		<p><ul class="digest">
+		<p><ul class="lig">
-			<li>1 µl Plasmid (pSB1C3-iBB39,pSB1A3-iBB49 and pSB1C3-iBB6315)</li>
+			<li>2 µl vector DNA (pSB1A3)</li>
-			<li>0.3 µl EcoRI</li>
+			<li>2 µl insert 1 DNA (iBB9)</li>
-			<li>0.3 µl PstI</li>
+			<li>2 µl insert 2 DNA (iBB10, iBB11, iBB12 or iBB13)</li>
-			<li>1 µl CutSmart</li>
+			<li>2 µl 10x T4 DNA ligase buffer</li>
-			<li>7.4 µl ddH2O</li>
+			<li>2 µl T4 DNA ligase</li>
+			<li>2 µl ddbidest. H2O</li>
 		</ul>
-		<p>The samples were incubated for 1 h at 37° C.</p>
+		<p>The samples were incubated overnight at room temperature.</p>
-		<table class="gel digest">
-			<colgroup>
-				<col width="50%" />
-				<col width="50%" />
-			</colgroup>
-		<thead>
-			<tr>
-				<th colspan="2" class="title">Gel electrophoresis</th>
-			</tr>
-		</thead>
-		<tbody>
-			<tr>
-				<td>
-					<img src="beispiel.png" width="50%" alt="gel-electrophoresis-image" />
-				</td>
-				<td>
-					<p>
-					<span class="gel-elc">Gel substances</span>
-					<ul class="gel-sub">
-						<li>1% Agarose gel</li>
-						<li>10 µl RedSafe in 50 ml gel</li>
-						<li>x µl Hyper Ladder</li>
-					</ul>
-					</p>
-					<p>
-						<span class="exp">Expactations</span>
-						<ul class="exp">
-							<li>Lane 1: 5300 kbp</li>
-							<li>Lane 2: 3300 kbp</li>
-							<li>Lane 3: 1300 kbp</li>
-						</ul>
-					</p>
-					<p><font color="#FF0000">After that, a gel electrophoresis was made, but failed and has to be repeated the next day</font></p>
-				</td>
-			</tr>
-		</tbody>
-		</table>
 	</div>
 </fieldset>
+</div>
-<fieldset class="experiment inoculation">
+<div class="notebooky-entry">
-    <legend><a name="ino">Inoculation</a></legend>
+<h2 class="title">
+	<a name="27-06-2013">27.06.2013</a>
+</h2>
+<fieldset class="experiment transformation">
+    <legend>Transformation</legend>
      <div class="investigator">
 		<span class="inv">Investigator:</span>
@@ Line 1,500: / Line 1,520: @@
      <div class="aim">
 		<span class="aim">Aim:</span>
-		<span class="aim-desc">inoculation of colonies for plasmid preparation</span>
+		<span class="aim-desc">Transformation of <i>E. coli</i> DH5&#945; with the plasmid DNA pSB1A3-iBB10+iBB9, pSB1A3-iBB11+iBB9, pSB1A3-iBB12+iBB9 and pSB1A3-iBB13+iBB9.</span>
 	</div>
 	<div class="exp-content">
-		<p>4 colonies of pSB1A3-iBB496315 were inoculated in 5 ml LB medium containing ampicillin.</p>
+		<p>The chemo competent <i>E. coli</i> DH5&alpha; cells were transformed with the plasmids pSB1A3-iBB10+iBB9, pSB1A3-iBB11+iBB9,pSB1A3-iBB12+iBB9 and pSB1A3-iBB13+iBB9 DNA and plated on LB-Amp-plates.<br />
+		The plates were incubated over night at 37 °C.</p>
 	</div>
 </fieldset>
-</div>
-<div class="notebooky-entry">
+<fieldset class="experiment inoculation">
-<h2 class="title">
+     <legend><a name="ino">Inoculation</a></legend>
-	<a name="02-07-2013">02.07.2013</a>
-</h2>
-<fieldset class="experiment miniprep">
-     <legend><a name="min">Miniprep</a></legend>
      <div class="investigator">
 		<span class="inv">Investigator:</span>
@@ Line 1,521: / Line 1,536: @@
      <div class="aim">
 		<span class="aim">Aim:</span>
-		<span class="aim-desc">preparation of the plasmid pSB1A3-iBB496315.</span>
+		<span class="aim-desc">inoculation of colonies for plasmid preparation</span>
 	</div>
 	<div class="exp-content">
-		<p>The plasmids were isolated with the “QIAprep Spin Miniprep Kit” (Qiagen, Düsseldorf) according to the producer instructions.</p>
+		<p>4 colonies of pSB1A3-iBB395146 and 4 colonies of pSB1A3-iBB496315 were inoculated in 5 ml LB medium containing ampicillin.</p>
 	</div>
 </fieldset>
@@ Line 1,532: / Line 1,547: @@
      <div class="investigator">
 		<span class="inv">Investigator:</span>
-		<span class="inv-names">Dominik</span>
+		<span class="inv-names">Alex</span>
 	</div>
      <div class="aim">
 		<span class="aim">Aim:</span>
-		<span class="aim-desc">Test digest of pSB1A3-iBB496315 and iBB4+iBB13+iBB96315 with <i>Eco</i>RI and <i>Pst</i>I</span>
+		<span class="aim-desc">Digest of pSB1A3-iBB395416 and pSB1A3-iBB496315 with <i>EcoR</i>I and <i>Pst</i>I.</span>
 	</div>
 	<div class="exp-content">
-		<p><ul class="digest">
+		<ul class="digest">
-			<li>1 µl Plasmid (pSB1A3-iBB496315 and iBB4+iBB13+iBB96315)</li>
+			<li>1 µl Plasmid DNA</li>
-			<li>0.3 µl EcoRI</li>
+			<li>0.3 µl <i>EcoR</i>I</li>
-			<li>0.3 µl PstI</li>
+			<li>0.3 µl <i>Pst</i>I</li>
-			<li>1 µl CutSmart</li>
+			<li>1 µl Cut Smart</li>
-			<li>7.4 µl ddH2O</li>
+			<li>7.4 µl ddbidest. H2O</li>
 		</ul>
-		<p>The samples were incubated for 1 h at 37° C.</p>
+		<p>The samples were incubated for 2 h at 37 °C.</p>
 		<table class="gel digest">
 			<colgroup>
@@ Line 1,560: / Line 1,575: @@
 			<tr>
 				<td>
-					<img src="beispiel.png" width="50%" alt="gel-electrophoresis-image" />
+					<img src="https://static.igem.org/mediawiki/2013/5/52/Gel_2013-06-27.png" width="100%" alt="gel-electrophoresis-image" />
 				</td>
 				<td>
@@ Line 1,567: / Line 1,582: @@
 					<ul class="gel-sub">
 						<li>1% Agarose gel</li>
-						<li>10 µl RedSafe in 50 ml gel</li>
+						<li>10 µl Ethidium bromide in 50 ml gel</li>
-						<li>x µl Hyper Ladder</li>
+						<li>4 µl 2-log DNA ladder (New England Biolabs)</li>
 					</ul>
 					</p>
 					<p>
-						<span class="exp">Expactations</span>
+							<table class="gel">
-						<ul class="exp">
+								<colgroup>
-							<li>Lane 1: 5300 kbp</li>
+									<col width="10%" />
-							<li>Lane 2: 3300 kbp</li>
+									<col width="2%" />
-							<li>Lane 3: 1300 kbp</li>
+									<col width="50%" />
-						</ul>
+									<col width="5%" />
+									<col width="33%" />
+								</colgroup>
+								<thead>
+									<th>Lane</th>
+									<th>&nbsp;</th>
+									<th>Content</th>
+									<th>&nbsp;</th>
+									<th>Expectations</th>
+								</thead>
+								<tr>
+									<td>upper:</td>
+								</tr>
+								<tr>
+									<td>2-5</td>
+									<th>&nbsp;</th>
+									<td>iBB395416 K1-4</td>
+									<th>&nbsp;</th>
+									<td>2400 + 2000 bp</td>
+								</tr>
+								<tr>
+									<td>6</td>
+									<th>&nbsp;</th>
+									<td>iBB39</td>
+									<th>&nbsp;</th>
+									<td>1000 + 2000 bp</td>
+								</tr>
+								<tr>
+								<tr>
+									<td>7</td>
+									<th>&nbsp;</th>
+									<td>iBB6416</td>
+									<th>&nbsp;</th>
+									<td>1400 + 2000 bp</td>
+								</tr>
+								<tr>
+									<td>lower:</td>
+								</tr>
+								<tr>
+									<td>2-6</td>
+									<th>&nbsp;</th>
+									<td>iBB496315 K1-5</td>
+									<th>&nbsp;</th>
+									<td>2400 + 2000 bp</td>
+								</tr>
+								<tr>
+									<td>7</td>
+									<th>&nbsp;</th>
+									<td>iBB49</td>
+									<th>&nbsp;</th>
+									<td>1200 + 2000 bp</td>
+								</tr>
+							</table>
 					</p>
-					<p>Worked as expected.</p>
+					<p>All clones show a double band above 2000 bp.</p>
+				</td>
+			</tr>
+			<tr>
+				<td colspan="2" class="gel-fazit">
+					<p>We received all expected fragments. The success of the ligation should be double-checked by means of PCR.</p>
 				</td>
 			</tr>
 		</tbody>
 		</table>
-	</div>
-</fieldset>
-</div>
-<div class="notebooky-entry">
-<h2 class="title">
-	<a name="14-08-2013">14.08.2013</a>
-</h2>
-<fieldset class="experiment sequencing">
-    <legend><a name="seq">Sequencing</a></legend>
-    <div class="investigator">
-		<span class="inv">Investigator:</span>
-		<span class="inv-names">Dominik</span>
-	</div>
-    <div class="aim">
-		<span class="aim">Aim:</span>
-		<span class="aim-desc">Examination of sequence analysis of the plasmids pSB1A3-iBB496315, pSB1A3-iBB4+iBB10+iBB96315 and pSB1A3-iBB4+iBB13+iBB96315.</span>
-	</div>
-	<div class="exp-content">
-		<p><ul class="seq">
-			<li>pSB1A3-iBB496315: Okay</li>
-			<li>pSB1A3-iBB4+iBB10+iBB96315: mutation in signal peptide that leads to Trp &rarr; Leu. Will be ignored</li>
-			<li>pSB1A3-iBB4+iBB13+iBB96315: mutation in terminator, will be ignored.</li>
-		</ul>
-		<p>All plasmids are brought to Marian who will transform them in <i>P. tricornutum</i> cells.</p>
 	</div>
 </fieldset>
@@ Line 1,615: / Line 1,661: @@
 </html>
-{{:Team:Marburg/Template:ContentEnd}}
+{{:Team:Marburg/Template:ContentEndNav}}
 {{:Team:Marburg/Template:Footer}}

Team:Marburg/Notebook:June

From 2013.igem.org

Latest revision as of 11:14, 27 October 2013

Notebook: June