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Team:DTU-Denmark/Notebook/8 August 2013
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! component(per reaction) !! without additives !! using 5% DMSO !! using 1M Betaine | ! component(per reaction) !! without additives !! using 5% DMSO !! using 1M Betaine | ||
Revision as of 08:09, 9 August 2013
8 August 2013
Contents |
lab 208
Main purpose
Who was in the lab
Kristian, Julia, Henrike
Procedure
PCR for AMO with USER endings
primers: 17a, 17b
template: gel purified AMO extraction fragment
Program: Standard with 54C annealing temperature and 3:00 extension time
Reaction Mix
| component(per reaction) | without additives | using 5% DMSO | using 1M Betaine |
|---|---|---|---|
| dNTPs | 1uL | 1uL | 1uL |
| HF buffer | 10uL | 10uL | 10uL |
| X7 polymerase | 0.5uL | 0.5uL | 0.5uL |
| MilliQ water | 31.5uL | 29uL | 21.5uL |
| template | 1uL | 1uL | 1uL |
| FW primer | 3uL | 3uL | 3uL |
| RV primer | 3uL | 3uL | 3uL |
| DMSO | - | 2.5uL | - |
| Betaine | - | - | 10uL |
PCR for extraction of Nir1
Results
Gel on yesterdays PCR
- 1kb ladder
- Yesterdays sample 1
- Yesterdays sample 2
- Yesterdays sample 3
- Yesterdays sample 4
- Neg.
- Neg.
- Nir2
- cycAX Histag
- 1kb ladder
Gel on ON PCR samples
- 1kb ladder
- Nir1
- Nir1 5%DMSO
- Nir1 1M Betaine
- Nir1 3M Betaine
- Nir2 3M Betaine U
- Nir2 1M Betaine U
- Nir2 5%DMSO U
- Nir2 U
- Neg (Nir)
- Ref 0%DMSO
- Ref 2%DMSO
- Ref 5%DMSO
- AMO U
- AMO 2%DMSO U
- Neg (AMO)
- NirG 5%DMSO
- NirG
- 1kb ladder
Gel on todays PCR samples
- 1kb ladder
- Nir Q5 poly
- Nir 2%DMSO Q5 poly
- Nir 3M Betaine Q5 poly
- Nir 5%DMSO Q5 poly
- Nir1 Q5 poly
- Nir1 2%DMSO Q5 poly
- Nir1 5%DMSO Q5 poly
- Nir1 3M Betaine Q5 poly
- AMO USER primers
- AMO +DMSO USER primers
- AMO +Betaine USER primers
- Nir1 DMSO
- Nir1 Betaine
- 1kb ladder
Conclusion
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