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Team:DTU-Denmark/Notebook/6 September 2013
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===Gel 3=== | ===Gel 3=== | ||
Revision as of 15:50, 6 September 2013
6 September 2013
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Contents |
Lab 208
Main purpose
- Add USER endings to HAO gene extracted from Nitrosomonas europea
Who was in the lab
Procedure
Repeat HAO USER PCR from yesterday with same conditions and programs
Template: HAO extraction fragment -isolated from Nitrosomonas europea- (purified from PCR run on 04.09)
Primers: 18a & 18b
Two versions, one with 5% DMSO and one with 3% DMSO and two programs: Touchdown (62C -> 55) and a single annealing temperature of 59C.
| compound | volume (uL) |
|---|---|
| dNTPs | 1 |
| HF | 10 |
| polymerase x7 | 0.5 |
| FW | 3 |
| RV | 3 |
| template | 1 |
| MQ | 29 |
| DMSO | 1.5 |
- Add 49 ul of the master mix, and
- Add 1 ul DMSO (for the 5%) or 1 ul of MQ (for the 3%)
Results
Gel 1
Gel of yesterday's PCR was empty (we lost the picture though...)
PCR was redone.
Gel 2
For the PCR results of today.
- 2-log ladder
- HAO User on 59C, 3% DMSO
- HAO User on 59C, 5% DMSO
- 2-log ladder
Gel 3
- 2-log ladder
- HAO User on touchdown, 3% DMSO
- HAO User on touchdown, 5% DMSO
- 2-log ladder
Conclusion
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