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Team:KIT-Kyoto/Notebook/Labnote
From 2014.igem.org
(Difference between revisions)
| Line 753: | Line 753: | ||
<tr> | <tr> | ||
<td>BL21(DE3)pLysS</td> | <td>BL21(DE3)pLysS</td> | ||
| - | <td colspan | + | <td colspan="2">pGEX6P-2</td> |
<td>non-induced</td> | <td>non-induced</td> | ||
</tr> | </tr> | ||
| Line 771: | Line 771: | ||
<br> | <br> | ||
<!--Protein Extraction(E.coli)--> | <!--Protein Extraction(E.coli)--> | ||
| - | <strong> | + | <strong>Protein Extraction(E.coli)</strong> |
<table class="materials"> | <table class="materials"> | ||
<tr> | <tr> | ||
| Line 800: | Line 800: | ||
<tr> | <tr> | ||
<td>BL21(DE3)pLysS</td> | <td>BL21(DE3)pLysS</td> | ||
| - | <td colspan | + | <td colspan="2">pGEX6P-2</td> |
<td>non-induced</td> | <td>non-induced</td> | ||
</tr> | </tr> | ||
| Line 818: | Line 818: | ||
<br> | <br> | ||
<!--Preparations for SDS-PAGE--> | <!--Preparations for SDS-PAGE--> | ||
| + | <strong>Preparations for SDS-PAGE</strong> | ||
Add SDS sample buffer to protein extractions in a flask, incubate at 37℃ for 15 minutes and keep it at 4℃. | Add SDS sample buffer to protein extractions in a flask, incubate at 37℃ for 15 minutes and keep it at 4℃. | ||
<!--Preparations for SDS-PAGE終わり--> | <!--Preparations for SDS-PAGE終わり--> | ||
Revision as of 08:15, 2 October 2014
LabNote
June
June 10, 2014
OKAMOTO
Miniprep
| Host | Sample | |
|---|---|---|
| Vector | Insert | |
| BL21(DE3) | pGEX6P-2 | CuMTSE1 |
| BL21(DE3) | pGEX6P-2 | CuMTSE2 |
| BL21(DE3) | pGEX6P-2 | CuMTS3 |
| BL21(DE3) | pGEX6P-2 | CuMTS62 |
Restriction Digest
| Host | Sample | Enzyme | Buffer | |
|---|---|---|---|---|
| Vector | Insert | |||
| BL21(DE3)PlysS | pGEX6P-2 | CuMTSE1 | EcoR1 | Buffer H |
| BL21(DE3)PlysS | pGEX6P-2 | CuMTSE2 | EcoR1 | Buffer H |
| BL21(DE3)PlysS | pGEX6P-2 | CuMTS3 | EcoR1 | Buffer H |
| BL21(DE3)PlysS | pGEX6P-2 | CuMTSE62 | EcoR1 | Buffer H |
| BL21(DE3)PlysS | pGEX6P-2 | CuMTSE1 | Sal1, Not1 | Buffer H |
| BL21(DE3)PlysS | pGEX6P-2 | CuMTSE2 | Sal1, Not1 | Buffer H |
| BL21(DE3)PlysS | pGEX6P-2 | CuMTS3 | Sal1, Not1 | Buffer H |
| BL21(DE3)PlysS | pGEX6P-2 | CuMTSE62 | Sal1, Not1 | Buffer H |
June 11, 2014
KOBAYASHI
AGE
| Lane | Sample/Marker | Enzyme | |
|---|---|---|---|
| Vector | Insert | ||
| 1 | λDNA-HindⅢ | ||
| 2 | pGEX6P-2 | CuMTSE1 | N.D |
| 3 | pGEX6P-2 | CuMTSE1 | EcoR1 |
| 4 | pGEX6P-2 | CuMTSE1 | Sal1,Not1 |
| 5 | pGEX6P-2 | CuMTSE2 | N.D |
| 6 | pGEX6P-2 | CuMTSE2 | EcoR1 |
| 7 | pGEX6P-2 | CuMTSE2 | Sal1, Not1 |
| 8 | pGEX6P-2 | CuMTS3 | N.D |
| 9 | pGEX6P-2 | CuMTS3 | EcoR1 |
| 10 | pGEX6P-2 | CuMTS62 | N.D |
| 11 | pGEX6P-2 | CuMTS62 | EcoR1 |
| 12 | pGEX6P-2 | CuMTS62 | Sal1,Not1 |
June 12, 2014
KOBAYASHI
Transformation
| Host | Sample | |
|---|---|---|
| Vector | Insert | |
| BL21(DE3)pLysS | pGEX6P-2 | CuMTSE1 |
| BL21(DE3)pLysS | pGEX6P-2 | CuMTSE2 |
| BL21(DE3)pLysS | pGEX6P-2 | CuMTS3 |
| BL21(DE3)pLysS | pGEX6P-2 | CuMTS62 |
June 13, 2014
KOBAYASHI
Colony Isolation
Select 6 colonies per plate from 4 plates from June 12th.
| Host | Sample | Section | |
|---|---|---|---|
| Vector | Insert | ||
| BL21(DE3)pLysS | pGEX6P-2 | CuMTSE1 | 1-a,1-b |
| BL21(DE3)pLysS | pGEX6P-2 | CuMTSE2 | 2-a,2-b |
| BL21(DE3)pLysS | pGEX6P-2 | CuMTS3 | 3-a,3-b |
| BL21(DE3)pLysS | pGEX6P-2 | CuMTSE62 | 6-a,6-b |
June 14, 2014
ONISHI
Miniprep
| Host | Sample | |
|---|---|---|
| Vector | Insert | |
| 1-a | pGEX6P-2 | CuMTSE1 |
| 1-b | pGEX6P-2 | CuMTSE1 |
| 2-a | pGEX6P-2 | CuMTSE2 |
| 2-b | pGEX6P-2 | CuMTSE2 |
| 3-a | pGEX6P-2 | CuMTS3 |
| 3-b | pGEX6P-2 | CuMTS3 |
| 6-a | pGEX6P-2 | CuMTS62 |
| 6-b | pGEX6P-2 | CuMTS62 |
Miniprep
| Host | Sample | |
|---|---|---|
| Vector | Insert | |
| BL21(DE3) | pGEX6P-2 | CuMTSE1 |
| BL21(DE3) | pGEX6P-2 | CuMTSE2 |
| BL21(DE3) | pGEX6P-2 | CuMTS3 |
| BL21(DE3) | pGEX6P-2 | CuMTS62 |
Restriction Digest
| Host | Sample | Enzyme | Buffer | |
|---|---|---|---|---|
| Vector | Insert | |||
| 1-a | pGEX6P-2 | CuMTSE1 | EcoR1 | Buffer H |
| 1-b | pGEX6P-2 | CuMTSE1 | EcoR1 | Buffer H |
| 2-a | pGEX6P-2 | CuMTSE2 | EcoR1 | Buffer H |
| 2-b | pGEX6P-2 | CuMTSE2 | EcoR1 | Buffer H |
| 3-a | pGEX6P-2 | CuMTS3 | EcoR1 | Buffer H |
| 3-b | pGEX6P-2 | CuMTS3 | EcoR1 | Buffer H |
| 6-a | pGEX6P-2 | CuMTS62 | EcoR1 | Buffer H |
| 6-b | pGEX6P-2 | CuMTS62 | EcoR1 | Buffer H |
| BL21(DE3) | pGEX6P-2 | CuMTSE1 | EcoR1 | Buffer H |
| BL21(DE3) | pGEX6P-2 | CuMTSE2 | EcoR1 | Buffer H |
| BL21(DE3) | pGEX6P-2 | CuMTS3 | EcoR1 | Buffer H |
| BL21(DE3) | pGEX6P-2 | CuMTS62 | EcoR1 | Buffer H |
AGE
| Lane | Sample/Marker | Insert | Enzyme | Buffer | |
|---|---|---|---|---|---|
| Host | Vector | ||||
| 1 | 1kb Ladder | ||||
| 2 | λDNA-HindⅢ | ||||
| 3 | 1-a | pGEX6P-2 | CuMTSE1 | EcoR1 | Buffer H |
| 4 | 1-b | pGEX6P-2 | CuMTSE1 | EcoR1 | Buffer H |
| 5 | 2-a | pGEX6P-2 | CuMTSE2 | EcoR1 | Buffer H |
| 6 | 2-b | pGEX6P-2 | CuMTSE2 | EcoR1 | Buffer H |
| 7 | 3-a | pGEX6P-2 | CuMTS3 | EcoR1 | Buffer H |
| 8 | 3-b | pGEX6P-2 | CuMTS3 | EcoR1 | Buffer H | 9 | 6-a | pGEX6P-2 | CuMTS62 | EcoR1 | Buffer H |
| 10 | 6-b | pGEX6P-2 | CuMTS62 | EcoR1 | Buffer H |
| 11 | BL21(DE3) | pGEX6P-2 | CuMTSE1 | EcoR1 | Buffer H |
| 12 | BL21(DE3) | pGEX6P-2 | CuMTSE2 | EcoR1 | Buffer H |
| 13 | BL21(DE3) | pGEX6P-2 | CuMTS3 | EcoR1 | Buffer H |
| 14 | BL21(DE3) | pGEX6P-2 | CuMTS62 | EcoR1 | Buffer H |
Applied λDNA-HindⅢ to compare and contrast AGE of July 14th and June 11th.
June 16, 2014
YAMAJI
Preculture
| Host | Sample | |
|---|---|---|
| Vector | Insert | |
| BL21(DE3)pLysS | pGEX6P-2 | CuMTSE1 |
| BL21(DE3)pLysS | pGEX6P-2 | CuMTSE2 |
| BL21(DE3)pLysS | pGEX6P-2 | CuMTS3 |
| BL21(DE3)pLysS | pGEX6P-2 | CuMTS62 |
| BL21(DE3)pLysS | pGEX6P-2 | |
Main Culture
| Host | Sample | IPTG | |
|---|---|---|---|
| Vector | Insert | ||
| BL21(DE3)pLysS | pGEX6P-2 | induced | |
| BL21(DE3)pLysS | pGEX6P-2 | CuMTSE1 | induced |
| BL21(DE3)pLysS | pGEX6P-2 | CuMTS2 | induced |
| BL21(DE3)pLysS | pGEX6P-2 | non-induced | |
| BL21(DE3)pLysS | pGEX6P-2 | CuMTS1 | non-induced |
| BL21(DE3)pLysS | pGEX6P-2 | CuMTSE2 | non-induced |
Protein Extraction(E.coli)
| Host | Sample | IPTG | |
|---|---|---|---|
| Vector | Insert | ||
| BL21(DE3)pLysS | pGEX6P-2 | induced | |
| BL21(DE3)pLysS | pGEX6P-2 | CuMTSE1 | induced |
| BL21(DE3)pLysS | pGEX6P-2 | CuMTS2 | induced |
| BL21(DE3)pLysS | pGEX6P-2 | non-induced | |
| BL21(DE3)pLysS | pGEX6P-2 | CuMTS1 | non-induced |
| BL21(DE3)pLysS | pGEX6P-2 | CuMTSE2 | non-induced |
Preparations for SDS-PAGE Add SDS sample buffer to protein extractions in a flask, incubate at 37℃ for 15 minutes and keep it at 4℃.
July
7/25
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August
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October
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