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Team:DTU-Denmark/Notebook/5 September 2013
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==Conclusion== | ==Conclusion== | ||
Revision as of 11:37, 6 September 2013
5 September 2013
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Contents |
Lab 208
Main purpose
PCR for HAO with USER endings after purification
Who was in the lab
Henrike, Ariadni
Procedure
PCR for HAO with USER endings
Template: HAO extraction fragment (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.
PCR reaction master mix (x5)
| 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 on yesterdays PCR
- 2-log ladder
- HAO USER on 59C, 3% DMSO
- HAO USER on 59C, 3% DMSO
- HAO USER on 57C, 3% DMSO
- HAO USER on 57C, 3% DMSO
- HAO USER on touchdown
- HAO USER on touchdown
- HAO USER on touchdown, 3% DMSO
- HAO USER on touchdown, 3% DMSO
- HAO extraction fragment on 57C, template -20 culture
- HAO extraction fragment on 57C, template -20 culture
- HAO extraction fragment on 57C, template -80 culture
- HAO extraction fragment on 57C, template -80 culture
- HAO extraction fragment on 59C, template -20 culture
- HAO extraction fragment on 59C, template -20 culture
- HAO extraction fragment on 59C, template -80 culture
- HAO extraction fragment on 59C, template -80 culture
- 2-log ladder
Conclusion
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