From 2013.igem.org

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Revision as of 10:13, 1 October 2013

Notebook: June

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
7.4 µl ddH2O

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 in 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.
Transformed cells were spread out on LB Amp culture plates.
Incubation ON at RT.
4 clones were picked for overnight 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 ddH2O

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

Expectations (upper gel)

Lane 2: 1,200 + 2,000 bp
Lane 3: 1,200 + 2,000 bp
Lane 4-7: 2,400 + 2,000 bp

Expectations (upper gel)

Lane 2: 1,500 + 2,000 bp
Lane 3: 1,000 + 2,000 bp
Lane 4-7: 2,400 + 2,000 bp

We don’t receive all expected fragments. The expected fragment in lane 1 is missing.

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	ddH2O

Gel electrophoresis

Gel substances

1% Agarose gel
10 µl Ethidium bromide 50 ml gel
4 µl 2-log DNA ladder

Expactations

Lane 1: ~ 6,000 bp for pSB1A3_486476315
Lane 2: 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 ddH2O

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

Gel electrophoresis

Gel substances

1% Agarose gel
10 µl RedSafe in 50 ml gel
x µl Hyper Ladder

Expactations

Lane 1: 5300 kbp
Lane 2: 3300 kbp
Lane 3: 1300 kbp

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 ddH2O

1 µl Plasmid (pSB1C3-iBB9)
0.5 µl XbaI
0.5 µl PstI
2 µl CutSmart
12 µl ddH2O

1 µl Plasmid (pSB1A3)
0.5 µl EcoRI
0.5 µl PstI
2 µl CutSmart
12 µl ddH2O

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

Gel electrophoresis

Gel substances

1% Agarose gel
10 µl RedSafe in 50 ml gel
x µl Hyper Ladder

Expactations

Lane 1: 5300 kbp
Lane 2: 3300 kbp
Lane 3: 1300 kbp

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

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 ddH2O

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.

02.07.2013

Miniprep

Investigator: Dominik

Aim: preparation of the plasmids pSB1A3-iBB10+iBB9, pSB1A3-iBB11+iBB9, pSB1A3-iBB12+iBB9 and pSB1A3-iBB13+iBB9.

The plasmids were isolated with the “QIAprep Spin Miniprep Kit” (Qiagen, Düsseldorf) according to the producer instructions.

Digest

Investigator: Dominik

Aim: Test digest of the plasmids pSB1A3-iBB10+iBB9, pSB1A3-iBB11+iBB9, pSB1A3-iBB12+iBB9 and pSB1A3-iBB13+iBB9 with EcoRI and PstI

1 µl Plasmid (pSB1A3-iBB10+iBB9, pSB1A3-iBB11+iBB9, pSB1A3-iBB12+iBB9 and pSB1A3-iBB13+iBB9)
0.3 µl EcoRI
0.3 µl PstI
1 µl CutSmart
7.4 µl ddH2O

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

Gel electrophoresis

Gel substances

1% Agarose gel
10 µl RedSafe in 50 ml gel
x µl Hyper Ladder

Expactations

Lane 1: 5300 kbp
Lane 2: 3300 kbp
Lane 3: 1300 kbp

Worked as expected.

23.07.2013

Digest

Investigator: Dominik

Aim: Test digest of the plasmids pSB1C3-iBB4+iBB11+iBB9, pSB1C3-iBB4+iBB12+iBB9 with EcoRI and PstI

1 µl Plasmid (pSB1C3-iBB4+iBB11+iBB9 and pSB1C3-iBB4+iBB12+iBB9)
0.3 µl EcoRI
0.3 µl PstI
1 µl CutSmart
7.4 µl ddH2O

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

Gel electrophoresis

Gel substances

1% Agarose gel
10 µl RedSafe in 50 ml gel
x µl Hyper Ladder

Expactations

Lane 1: 5300 kbp
Lane 2: 3300 kbp
Lane 3: 1300 kbp

Worked as expected.

24.07.2013

Inoculation

Investigator: Dominik

Aim: inoculation of colonies for plasmid preparation

4 colonies of pSB1C3-iBB4+iBB10+iBB9 and 4 colonies of pSB1C3-iBB4+iBB13+iBB9 were inoculated in 5 ml LB medium containing chloramphenicol.

Digest

Investigator: Dominik

Aim: Digest of pSB1C3-iBB4+iBB11+iBB9, pSB1C3-iBB4+iBB12+iBB9, pSB1C3-iBB6315 and pSB1A3.

1000 ng Plasmid (pSB1C3-iBB4+iBB11+iBB9, pSB1C3-iBB4+iBB12+iBB9)
0.5 µl EcoRI
0.5 µl SpeI
2 µl CutSmart
ad 20 µl ddH2O

1000 ng Plasmid (pSB1C3-iBB6315)
0.5 µl XbaI
0.5 µl PstI
2 µl CutSmart
ad 20 µl ddH2O

1000 ng Plasmid (pSB1A3)
0.5 µl EcoRI
0.5 µl PstI
2 µl CutSmart
ad 20 µl ddH2O

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

Gel electrophoresis

Gel substances

1% Agarose gel
10 µl RedSafe in 50 ml gel
x µl Hyper Ladder

Expactations

Lane 1: 5300 kbp
Lane 2: 3300 kbp
Lane 3: 1300 kbp

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

Ligation

Investigator: Dominik

Aim: Assemble the biobricks iBB4+iBB11+iBB96315 and iBB4+iBB12+iBB96315 into the vector pSB1A3.

2 µl vector DNA (pSB1A3)
5 µl insert 1 DNA (iBB6315)
5 µl insert 2 DNA (iBB4+iBB11+iBB9, iBB4+iBB12+iBB9)
2 µl 10x T4 DNA ligase buffer
2 µl T4 DNA ligase
4 µl ddH2O

The samples were incubated for 2h at 16 °C.

Transformation

Investigator: Dominik

Aim: Transformation of E. coli DH5α with the plasmid DNA pSB1A3-iBB4+iBB11+iBB96315 and pSB1A3-iBB4+iBB12+iBB96315.

The chemo competent E. coli DH5α cells were transformed with the plasmids pSB1A3-iBB4+iBB11+iBB96315 and pSB1A3-iBB4+iBB12+iBB96315 DNA and plated on LB-Amp-plates.
The plates were incubated over night at 37° C.

25.07.2013

Sequencing

Investigator: Dominik

Aim: Complete sequencing of pSB1A3-iBB496315.

8 new sequencing samples were sent out.

Miniprep

Investigator: Dominik

Aim: preparation of the plasmids pSB1A3-iBB4+iBB10+iBB9 and pSB1A3-iBB4+iBB13+iBB9.

The plasmids were isolated with the “QIAprep Spin Miniprep Kit” (Qiagen, Düsseldorf) according to the producer instructions.

Digest

Investigator: Dominik

Aim: Test digest of pSB1C3-iBB4+iBB10+iBB9 and pSB1C3-iBB4+iBB13+iBB9 with EcoRI and PstI

1 µl Plasmid (pSB1C3-iBB4+iBB10+iBB9 and pSB1C3-iBB4+iBB13+iBB9)
0.3 µl EcoRI
0.3 µl PstI
1 µl CutSmart
7.4 µl ddH2O

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

Gel electrophoresis

Gel substances

1% Agarose gel
10 µl RedSafe in 50 ml gel
x µl Hyper Ladder

Expactations

Lane 1: 5300 kbp
Lane 2: 3300 kbp
Lane 3: 1300 kbp

Two samples of each plasmid preparation showed the expected fragments.

Inoculation

Investigator: Dominik

Aim: inoculation of colonies for plasmid preparation

4 colonies of pSB1A3-iBB4+iBB11+iBB96315 and 4 colonies of pSB1A3-iBB4+iBB12+iBB96315 were inoculated in 5 ml LB medium containing chloramphenicol.

Competent cells

Investigator: Dominik

Aim: Overnight culture for making new aliquots of competent cells.

For making new aliquots of E. coli DH5α cells 50 ml LB medium were inoculated with 500 µl of an E. coli DH5α culture.

26.07.2013

Digest

Investigator: Dominik

Aim: Digest of pSB1C3-iBB4+iBB10+iBB9, pSB1C3-iBB4+iBB13+iBB9, pSB1C3-iBB6315 and pSB1A3.

1000 ng Plasmid (pSB1C3-iBB4+iBB10+iBB9, pSB1C3-iBB4+iBB13+iBB9)
0.5 µl EcoRI
0.5 µl SpeI
2 µl CutSmart
ad 20 µl ddH2O

1000 ng Plasmid (pSB1C3-iBB6315)
0.5 µl XbaI
0.5 µl PstI
2 µl CutSmart
ad 20 µl ddH2O

1000 ng Plasmid (pSB1A3)
0.5 µl EcoRI
0.5 µl PstI
2 µl CutSmart
ad 20 µl ddH2O

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

Gel electrophoresis

Gel substances

1% Agarose gel
10 µl RedSafe in 50 ml gel
x µl Hyper Ladder

Expactations

Lane 1: 5300 kbp
Lane 2: 3300 kbp
Lane 3: 1300 kbp

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

Ligation

Investigator: Dominik

Aim: Assemble the biobricks iBB4+iBB10+iBB9, iBB4+iBB13+iBB9 and iBB96315 into the vector pSB1A3.

2 µl vector DNA (pSB1A3)
5 µl insert 1 DNA (iBB96315)
5 µl insert 2 DNA (iBB4+iBB10+iBB9,iBB4+iBB13+iBB9)
2 µl 10x T4 DNA ligase buffer
2 µl T4 DNA ligase
4 µl ddH2O

The samples were incubated for 1h at 16 °C.

Resulting in the plasmids pSB1A3-iBB4+iBB10+iBB96315 and pSB1A3-iBB4+iBB13+iBB96315.

Miniprep

Investigator: Dominik

Aim: preparation of the pSB1A3-iBB4+iBB11+iBB96315 and pSB1A3-iBB4+iBB12+iBB96315.

The plasmids were isolated with the “QIAprep Spin Miniprep Kit” (Qiagen, Düsseldorf) according to the producer instructions.

Digest

Investigator: Dominik

Aim: Test digest of ppSB1A3-iBB4+iBB11+iBB96315 and pSB1A3-iBB4+iBB12+iBB96315 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 ddH2O

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

Gel electrophoresis

Gel substances

1% Agarose gel
10 µl RedSafe in 50 ml gel
x µl Hyper Ladder

Expactations

Lane 1: 5300 kbp
Lane 2: 3300 kbp
Lane 3: 1300 kbp

Most of the preparations resulted in the expected fragments, all others were discarded.

Competent cells

Investigator: Dominik

Aim: New aliquots of competent cells (E. coli DH5α).

About 80 new aliquots were made.

02.07.2013

Sequencing

Investigator: Dominik

Aim: Analysis of the sequence of pSB1A3-iBB496315.

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.

Transformation

Investigator: Dominik

Aim: Transformation of E. coli DH5α with the plasmid DNA pSB1A3-iBB4+iBB10+iBB96315 and pSB1A3-iBB4+iBB13+iBB96315.

The chemo competent E. coli DH5α cells were transformed with the plasmids pSB1A3-iBB4+iBB10+iBB96315 and pSB1A3-iBB4+iBB13+iBB96315 and plated on LB-Amp-plates.
The plates were incubated over night at 37° C.

30.07.2013

Digest

Investigator: Dominik

Aim: Digest of pSB1A3-iBB49 and pSB1C3-iBB6315.

1000 ng Plasmid (pSB1A3-iBB49)
0.5 µl PstI
0.5 µl SpeI
2 µl CutSmart
ad 20 µl ddH2O

1000 ng Plasmid (pSB1C3-iBB6315)
0.5 µl XbaI
0.5 µl PstI
2 µl CutSmart
ad 20 µl ddH2O

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

Gel electrophoresis

Gel substances

1% Agarose gel
10 µl RedSafe in 50 ml gel
x µl Hyper Ladder

Expactations

Lane 1: 5300 kbp
Lane 2: 3300 kbp
Lane 3: 1300 kbp

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

Inoculation

Investigator: Dominik

Aim: inoculation of colonies for plasmid preparation

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. Additionally the remaining ligation preparation was transformed in E. coli DH5α?????.

31.07.2013

Ligation

Investigator: Dominik

Aim: Ligation of pSB1A3-iBB49 and iBB6315.

2 µl vector DNA (pSB1A3-iBB49) (40 ng)
5 µl insert DNA (iBB6315) (150 ng)
2 µl 10x T4 DNA ligase buffer
2 µl T4 DNA ligase
9 µl ddH2O

The samples were incubated for 1h at 16 °C.

Transformation

Investigator: Dominik

Aim: Transformation of E. coli DH5α with the plasmid DNA pSB1A3-iBB49+iBB6315.

The chemo competent E. coli DH5α cells were transformed with the plasmid pSB1A3-iBB49+iBB6315 and plated on LB-Amp-plates.
The plates were incubated over night at 37° C.

Miniprep

Investigator: Dominik

Aim: Preparation of the pSB1A3-iBB4+iBB10+iBB96315 and pSB1A3-iBB4+iBB13+iBB96315.

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.

Digest

Investigator: Dominik

Aim: Test digest of pSB1A3-iBB4+iBB10+iBB96315 with EcoRI and PstI

1 µl Plasmid (pSB1A3-iBB4+iBB10+iBB96315)
0.3 µl EcoRI
0.3 µl PstI
1 µl CutSmart
7.4 µl ddH2O

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

Gel electrophoresis

Gel substances

1% Agarose gel
10 µl RedSafe in 50 ml gel
x µl Hyper Ladder

Expactations

Lane 1: 5300 kbp
Lane 2: 3300 kbp
Lane 3: 1300 kbp

Worked as expected.

Sequencing

Investigator: Dominik

Aim: Examination of the sequence of pSB1A3-iBB4+iBB10+iBB96315.

8 samples were sent out for sequencing.

Inoculation

Investigator: Dominik

Aim: inoculation of colonies for plasmid preparation

Both colonies of pSB1A3-iBB4+iBB13+iBB96315 were inoculated in 5 ml LB medium containing ampicillin.

01.08.2013

Sequencing

Investigator: Dominik

Aim: Examination of the sequence of pSB1A3-iBB4+iBB10+iBB96315.

2 additional sequence samples were sent out for sequencing.

Digest

Investigator: Dominik

Aim: Test digest of pSB1A3-iBB4+iBB13+iBB96315 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 ddH2O

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

Gel electrophoresis

Gel substances

1% Agarose gel
10 µl RedSafe in 50 ml gel
x µl Hyper Ladder

Expactations

Lane 1: 5300 kbp
Lane 2: 3300 kbp
Lane 3: 1300 kbp

After that, a gel electrophoresis was made, but failed and has to be repeated the next day

Inoculation

Investigator: Dominik

Aim: inoculation of colonies for plasmid preparation

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

02.07.2013

Miniprep

Investigator: Dominik

Aim: preparation of the plasmid pSB1A3-iBB496315.

The plasmids were isolated with the “QIAprep Spin Miniprep Kit” (Qiagen, Düsseldorf) according to the producer instructions.

Digest

Investigator: Dominik

Aim: Test digest of pSB1A3-iBB496315 and iBB4+iBB13+iBB96315 with EcoRI and PstI

1 µl Plasmid (pSB1A3-iBB496315 and iBB4+iBB13+iBB96315)
0.3 µl EcoRI
0.3 µl PstI
1 µl CutSmart
7.4 µl ddH2O

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

Gel electrophoresis

Gel substances

1% Agarose gel
10 µl RedSafe in 50 ml gel
x µl Hyper Ladder

Expactations

Lane 1: 5300 kbp
Lane 2: 3300 kbp
Lane 3: 1300 kbp

Worked as expected.

14.08.2013

Sequencing

Investigator: Dominik

Aim: Examination of sequence analysis of the plasmids pSB1A3-iBB496315, pSB1A3-iBB4+iBB10+iBB96315 and pSB1A3-iBB4+iBB13+iBB96315.

pSB1A3-iBB496315: Okay
pSB1A3-iBB4+iBB10+iBB96315: mutation in signal peptide that leads to Trp → Leu. Will be ignored
pSB1A3-iBB4+iBB13+iBB96315: mutation in terminator, will be ignored.

All plasmids are brought to Marian who will transform them in P. tricornutum cells.

@@ Line 30: / Line 30: @@
 		</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>
+                 <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 ligation">
+    <legend><a name="lig">Ligation, Transformation, Plasmid prep</a></legend>
+    <div class="investigator">
+		<span class="inv">Investigator:</span>
+		<span class="inv-names">Alex</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>
+	</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>Ligation product were transformed into chemical competent DH5α cells.</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 cultures.</li>
+                        <li>A plasmid preparation was carried out the next day.</li>
+		</ul>
+	</div>
+</fieldset>
+</div>
+<div class="notebooky-entry">
+<h2 class="title">
+	<a name="22-06-2013">22.06.2013</a>
+</h2>
+<fieldset class="experiment digest">
+    <legend><a name="dig">Test digest</a></legend>
+    <div class="investigator">
+		<span class="inv">Investigator:</span>
+		<span class="inv-names">Alex</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>
+	</div>
+	<div class="exp-content">
+		<ul class="digest">
+			<li>1 µl Plasmid DNA</li>
+			<li>0.3 µl EcoRI</li>
+			<li>0.3 µL PstI</li>
+			<li>1 µl Cut Smart</li>
+			<li>7.4 µl ddH2O</li>
+		</ul>
+		<p>The samples were incubated for 2 h at 37° C.</p>
 		<table class="gel digest">
 			<colgroup>
@@ Line 51: / Line 101: @@
 					<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</li>
 					</ul>
 					</p>
 					<p>
-						<span class="exp">Expactations</span>
+						<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>
+                                                <span class="exp">Expectations (upper gel)</span>
 						<ul class="exp">
-							<li>Lane 1: 5300 kbp</li>
+							<li>Lane 2: 1,500 + 2,000 bp</li>
-							<li>Lane 2: 3300 kbp</li>
+							<li>Lane 3: 1,000 + 2,000 bp</li>
-							<li>Lane 3: 1300 kbp</li>
+							<li>Lane 4-7: 2,400 + 2,000 bp</li>
 						</ul>
 					</p>
@@ Line 75: / Line 131: @@
 	</div>
 </fieldset>
 </div>

Team:Marburg/Notebook:June

From 2013.igem.org

Revision as of 10:13, 1 October 2013

Notebook: June