Team:Kyoto/Notebook

From 2013.igem.org

(Difference between revisions)
(→Aug 21)
(→Ristriction Enzyme Proccessing)
Line 297: Line 297:
</div>  
</div>  
====Ristriction Enzyme Proccessing====
====Ristriction Enzyme Proccessing====
 +
<div class="experiment">
 +
<span class="author">by Kojima and Nakamoto</span>
 +
{| class="wikitable"
 +
!8/20 Ptet-(1) ||EcoRI||XbaI||10x BSA||10x buffer M||MilliQ||total
 +
|-
 +
|4.5||1||1||3||3||17.5||30
 +
|-
 +
|0.5||0.2||0||1||1||7.3||10
 +
|-
 +
|0.5||0||0.2||1||1||7.3||10
 +
|-
 +
|0.5||0||0||1||1||7.5||10
 +
|}
 +
 +
{| class="wikitable"
 +
!8/20 Pconst-RBS-luxR-DT-(2) ||EcoRI||XbaI||10x BSA||10x buffer M||MilliQ||total
 +
|-
 +
|2.9||1||1||3||3||19.1||30
 +
|-
 +
|0.3||0.2||0||1||1||7.5||10
 +
|-
 +
|0.3||0||0.2||1||1||7.5||10
 +
|-
 +
|0.3||0||0||1||1||7.7||10
 +
|}
 +
 +
{| class="wikitable"
 +
!8/20 Ptet-(1) ||EcoRI||XbaI||10x BSA||10x buffer M||MilliQ||total
 +
|-
 +
|4.5||1||1||3||3||17.5||30
 +
|-
 +
|0.5||0.2||0||1||1||7.3||10
 +
|-
 +
|0.5||0||0.2||1||1||7.3||10
 +
|-
 +
|0.5||0||0||1||1||7.5||10
 +
|}
 +
</div>

Revision as of 03:57, 23 August 2013

Home Team Official Team Profile Project Parts Submitted to the Registry Modeling Notebook Material and method Safety Attributions


Contents

template

transformation

じっけんしゃ

NameWellSampleCompetent CellsTotalPlate
いでんしぱーつのうぇるなんちゃらµLごにょごにょµLほにゃららµLこーせーぶっしつ
いかどうよう

Aug 20

Miniprep

by kojima

DNAconcentration[µg/µL]260/280260/230
8/18 Ptet -12201.691.87
8/18 Ptet -22101.691.71
8/18 RBS-tetR-DT -12361.631.69
8/18 RBS-tetR-DT -21821.651.98
8/18 J23100-RBS-GFP-DT -13341.712.12
8/18 J23100-RBS-luxR-DT -14401.671.60
8/18 J23100-RBS-luxR-DT -23441.681.83
8/18 J23100-RBS-lacZα-DT -12801.701.81
8/18 RBS-lysis3 -12821.671.76
8/18 RBS-lysis3 -22121.301.35

LB Culture Medium

by Okazaki

volume200mL
Bacto Trypton2g
Bacto yeast extract1g
Nacl1g
Agar Powder2g
Amp40µl

1.Measuring reagents and put they in the Erlenmeyer flask
2.diluting in measuring cylinder to 200ml total
3.autoclaving
4.adding ampicirine 40ml


not written yet

Ristriction Enzyme Proccessing

by Kojima and Ashida

8/20 Ptet -1(220µg/mL)EcoRIXbaI10x BSA10x buffer MMilliQtotal
4.5113317.530
0.50.20117.310
0.500.2117.310
0.500117.510
8/20 RBS-tetR-DT -2(182µg/mL)EcoRISpeI10x buffer HMilliQtotal
5.511319.530
0.60.2018.210
0.600.218.210
0.60018.410
8/20 J23100-RBS-GFP-DT(334µg/ml)EcoRISpeI10x buffer HMilliQtotal
3.01132230
0.30.2018.510
0.300.218.510
0.30018.710
8/20 J23100-RBS-luxR-DT -2(344µg/mL)EcoRIXbaIBSA10x buffer MMilliQtotal
2.9113319.130
0.30.20117.510
0.300.2117.510
0.300117.710
8/20 RBS-lysis3 -1(282µg/mL)EcoRISpeI10x buffer HMilliQtotal
3.511321.530
0.40.2018.410
0.400.218.410
0.40018.610
8/17 DT -1(188µg/mL)EcoRIXbaI10x BSA10x buffer MMilliQtotal
5.3113316.730
0.30.20117.510
0.300.2117.510
0.300117.710

incubate ~20:00

Transformation

by Nakamoto

NameSampleCompetent Cells(XL10-gold)TotalPlate
tRNA-Spinach-tRNA1µL10µL11µL8/19 LB+Amp
tetR-aptamer 12_PC(076)1µL10µL11µL8/19 LB+Amp
tetR-aptamer 12_1R(113)1µL10µL11µL8/19 LB+Amp
tetR-aptamer 12_1M(105)1µL10µL11µL8/19 LB+Amp
pT181 attenuator1µL10µL11µL8/19 LB+Amp
Fusin1 attenuator1µL10µL11µL8/19 LB+Amp
Fusion3m2 attenuator1µL10µL11µL8/19 LB+Amp
pT181 antisense1µL10µL11µL8/19 LB+Amp
Fusion1 antisense1µL10µL11µL8/19 LB+Amp
Fusion6 antisense1µL10µL11µL8/19 LB+Amp

incubate 37°C overnight 20:34~

Electrophoresis

by Kojima

LaneSampleEnzyme1Enzyme2
18/20 Ptet -1EcoRIXbaI
28/20 Ptet -1EcoRI-
38/20 Ptet -1-XbaI
48/20 Ptet -1--
58/20 RBS-tetR-DT -2EcoRISpeI
68/20 RBS-tetR-DT -2EcoRI-
78/20 RBS-tetR-DT -2-SpeI
88/20 RBS-tetR-DT -2--
91kbp ladder--
108/20 J23100-RBS-GFP-DT -1EcoRISpeI
118/20 J23100-RBS-GFP-DT -1EcoRI-
128/20 J23100-RBS-GFP-DT -1-SpeI
138/20 J23100-RBS-GFP-DT -1--
148/20 J23100-RBS-luxR-DT -1EcoRIXbaI
158/20 J23100-RBS-luxR-DT -1EcoRI-
168/20 J23100-RBS-luxR-DT -1-XbaI
178/20 J23100-RBS-luxR-DT -1--
181kbp ladder--
198/20 RBS-lysis3 -1EcoRISpeI
208/20 RBS-lysis3 -1EcoRI-
218/20 RBS-lysis3 -1-SpeI
228/20 RBS-lysis3 -1--
231kbp ladder--
248/17 DTEcoRIXbaI
258/17 DTEcoRI-
268/17 DT-XbaI
278/17 DT--

Aug 21

Plusgrow medium(+amp)

by Tatsui

  • material
  • 8/17 Plusgrow medium 50ml
  • 7/22 5000×　Ampicillin 10µl


  1. Measuring materials and putting them in a 50ml tube

Liquid culture

by tatsui

Samplemedium
8/20 tRNA-spinach8/21 Plusgrow medium(+amp)
8/20 TetR-aptamer 12_P(1076)8/21 Plusgrow medium(+amp)
8/20 TetR-aptamer 12_1R(113)8/21 Plusgrow medium(+amp)
8/20 TetR-aptamer 12_1M(105) 8/21 Plusgrow medium(+amp)
8/20 PT181 attenuator8/21 Plusgrow medium(+amp)
8/20 Fusion1 attenuator 8/21 Plusgrow medium(+amp)
8/20 Fusion3m2 attenuator 8/21 Plusgrow medium(+amp)
8/20 PT181 antisense 8/21 Plusgrow medium(+amp)
8/20 Fusion1 antisense 8/21 Plusgrow medium(+amp)
8/20 Fusion6 antisense 8/21 Plusgrow medium(+amp)

incubate 37°C 10hour

Ristriction Enzyme Proccessing

by Kojima and Nakamoto

8/20 Ptet-(1) EcoRIXbaI10x BSA10x buffer MMilliQtotal
4.5113317.530
0.50.20117.310
0.500.2117.310
0.500117.510
8/20 Pconst-RBS-luxR-DT-(2) EcoRIXbaI10x BSA10x buffer MMilliQtotal
2.9113319.130
0.30.20117.510
0.300.2117.510
0.300117.710
8/20 Ptet-(1) EcoRIXbaI10x BSA10x buffer MMilliQtotal
4.5113317.530
0.50.20117.310
0.500.2117.310
0.500117.510
Retrieved from "http://2013.igem.org/Team:Kyoto/Notebook"