Team:Kyoto/Notebook

From 2013.igem.org

(Difference between revisions)
m
m
Line 52: Line 52:
===Miniprep===
===Miniprep===
<div class="experiment">
<div class="experiment">
-
<span class="author">by kojima </span>
+
<span class="author">Kojima </span>
{|class="wikitable"  
{|class="wikitable"  
!DNA!!concentration[&micro;g/&micro;L]!!260/280!!260/230  
!DNA!!concentration[&micro;g/&micro;L]!!260/280!!260/230  
Line 79: Line 79:
===LB Culture Medium===
===LB Culture Medium===
<div class="experiment">
<div class="experiment">
-
<span class="author">by Okazaki</span>
+
<span class="author">Okazaki</span>
{|class="wikitable"  
{|class="wikitable"  
!volume||200mL
!volume||200mL
Line 104: Line 104:
===Ristriction Enzyme Proccessing===
===Ristriction Enzyme Proccessing===
<div class="experiment">
<div class="experiment">
-
<span class="author">by Kojima and Ashida</span>
+
<span class="author">Kojima and Ashida</span>
{|class="wikitable"  
{|class="wikitable"  
!8/20 Ptet -1(220&micro;g/mL)!!EcoRI!!XbaI!!10x BSA!!10x buffer M!!MilliQ!!total
!8/20 Ptet -1(220&micro;g/mL)!!EcoRI!!XbaI!!10x BSA!!10x buffer M!!MilliQ!!total
Line 176: Line 176:
===Transformation===
===Transformation===
<div class="experiment">
<div class="experiment">
-
<span class="author">by Nakamoto</span>
+
<span class="author">Nakamoto</span>
{| class="wikitable"
{| class="wikitable"
!Name||Sample||Competent Cells(XL10-gold)||Total||Plate
!Name||Sample||Competent Cells(XL10-gold)||Total||Plate
Line 205: Line 205:
===Electrophoresis===
===Electrophoresis===
<div class="experiment">
<div class="experiment">
-
<span class="author">by Kojima</span>
+
<span class="author">Kojima</span>
{| class="wikitable"
{| class="wikitable"
!Lane||Sample||Enzyme1||Enzyme2
!Lane||Sample||Enzyme1||Enzyme2
Line 268: Line 268:
====Plusgrow medium(+amp)====
====Plusgrow medium(+amp)====
<div class="experiment">
<div class="experiment">
-
<span class="author">by Tatsui</span>
+
<span class="author">Tatsui</span>
*material
*material
:*8/17 Plusgrow medium 50ml
:*8/17 Plusgrow medium 50ml
Line 278: Line 278:
====Liquid culture====
====Liquid culture====
<div class="experiment">
<div class="experiment">
-
<span class="author">by tatsui</span>
+
<span class="author">Tatsui</span>
{| class="wikitable"
{| class="wikitable"
!Sample||medium
!Sample||medium
Line 308: Line 308:
====Ristriction Enzyme Proccessing====
====Ristriction Enzyme Proccessing====
<div class="experiment">
<div class="experiment">
-
<span class="author">by Kojima and Nakamoto</span>
+
<span class="author">Kojima and Nakamoto</span>
{| class="wikitable"
{| class="wikitable"
!8/20 Ptet-(1) ||EcoRI||XbaI||10x BSA||10x buffer M||MilliQ||total
!8/20 Ptet-(1) ||EcoRI||XbaI||10x BSA||10x buffer M||MilliQ||total
Line 354: Line 354:
===Liquid culture===
===Liquid culture===
<div class="experiment">
<div class="experiment">
-
<span class="author">by Nakamoto</span>
+
<span class="author">Nakamoto</span>
{| class="wikitable"
{| class="wikitable"
!|Sample||Medium
!|Sample||Medium
Line 393: Line 393:
===colony PCR===
===colony PCR===
<div class="experiment">
<div class="experiment">
-
<span class="author">by Kojima</span>
+
<span class="author">Kojima</span>
{| class="wikitable"
{| class="wikitable"
!Sample||base pair
!Sample||base pair

Revision as of 06:40, 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 19

Aug 20

Miniprep

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

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

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

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

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)

Tatsui

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


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

Liquid culture

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

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
8/17 DTEcoRIXbaI10x BSA10x buffer MMilliQtotal
5.3113316.730
0.50.20117.310
0.500.2117.310
0.500117.510


incubate 37°C 1h

Electrophoresis

Aug 22

Liquid culture

Nakamoto

SampleMedium
8/21 Pcon-RBS-GFP-DT-Pcon-RBS-luxR-DT (1)8/21 Plusgrow medium(+amp)
8/21 Pcon-RBS-GFP-DT-Pcon-RBS-luxR-DT (2)8/21 Plusgrow medium(+amp)
8/21 Pcon-RBS-GFP-DT control (1)8/21 Plusgrow medium(+amp)
8/21 Pcon-RBS-GFP-DT control (2)8/21 Plusgrow medium(+amp)
8/21 RBS-lysis1 (1)8/21 Plusgrow medium(+amp)
8/21 RBS-lysis1 (2)8/21 Plusgrow medium(+amp)
8/21 RBS control (1)8/21 Plusgrow medium(+amp)
8/21 RBS control (2)8/21 Plusgrow medium(+amp)
8/21 Ptet(pm)-RBS-tetR-DT (1)8/21 Plusgrow medium(+amp)
8/21 Ptet(pm)-RBS-tetR-DT (2)8/21 Plusgrow medium(+amp)
8/21 Ptet(pm) control (1)8/21 Plusgrow medium(+amp)
8/21 Ptet(pm) control8/21 Plusgrow medium(+amp)
8/21 Plux-RBS-GFP-DT (1)8/21 Plusgrow medium(+amp)
8/21 pSB1C3(BBa_J04450) (1)8/21 Plusgrow medium(+amp)
8/21 pSB1C3(BBa_J04450) (2)8/21 Plusgrow medium(+amp)

incubate 37°C 10hour

colony PCR

Kojima

Samplebase pair
8/21 RBS-lysis1(1)400
8/21 RBS-lysis1(2)400
8/21 RBS control(1)
8/21 RBS control(2)
negative control
Samplebase pair
8/21 Pcon-GFP-DT-Pcon-RBS-luxR-DT (1)2143
8/21 Pcon-GFP-DT-Pcon-RBS-luxR-DT (2)2143
8/21 Pcon-RBS-luxR control(1)
8/21 Pcon-RBS-luxR control(2)
negative control
Samplebase pair
8/21 Ptet(pm)-RBS-tetR-DT (1)
8/21 Ptet(pm)-RBS-tetR-DT (2)
8/21 Ptet(pm) control (1)
8/21 Ptet(pm) control (2)
8/21 Plux-RBS-GFP-DT (1)
8/21 pSB1C3(BBa_J04450) (1)
8/21 pSB1C3(BBa_J04450) (2)
negative control
Retrieved from "http://2013.igem.org/Team:Kyoto/Notebook"