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Team:Kyoto/Notebook1
From 2012.igem.org
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After 42°C 60sec heat shock, cells were stored on ice for 2min.<br> | After 42°C 60sec heat shock, cells were stored on ice for 2min.<br> | ||
Then cells were preincubated at 37°C for 1hr, plated to Kanamycin plate. | Then cells were preincubated at 37°C for 1hr, plated to Kanamycin plate. | ||
| + | <br> | ||
| + | <br> | ||
| + | ==August 13== | ||
| + | Liquid culture of FT | ||
| + | <br> | ||
| + | <br> | ||
| + | ==August 14== | ||
Latest revision as of 13:24, 21 August 2012
August 2
Mutation of FT
| 10xBufer | 2mM dNTPs | primer fwd | primer rev | template | polymerase | MilliQ | Total |
|---|---|---|---|---|---|---|---|
| 5 | 5 | 1.5 | 1.5 | 0.5 | 1 | 35.5 | 50 |
94°C 2min, (98°C 10sec, 68°C 4min)x2cycles, 4°C Hold
| PCR product | Dpn1 |
|---|---|
| 50 | 2 |
| product | MilliQ | Ligase | T4 Kinase | Total |
|---|---|---|---|---|
| 2 | 7 | 5 | 1 | 15 |
16°C, 1h incubate
| competent cell | DNA |
|---|---|
| 20 | 2 |
Cells were stored on ice for 30min.
After 42°C 60sec heat shock, cells were stored on ice for 2min.
Then cells were preincubated at 37°C for 1hr, plated to Kanamycin plate.
August 13
Liquid culture of FT
