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Team:DTU-Denmark/Notebook/4 August 2013
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| - | =lab | + | =lab 208= |
<hr/> | <hr/> | ||
| + | |||
==Main purpose== | ==Main purpose== | ||
<hr/> | <hr/> | ||
| + | Amplify the two Nir fragments and prepare for PCR with USER primers. | ||
==Who was in the lab== | ==Who was in the lab== | ||
<hr/> | <hr/> | ||
| + | Kristian | ||
==Procedure== | ==Procedure== | ||
<hr/> | <hr/> | ||
| + | |||
| + | ===PCR for amplification of Nir fragments=== | ||
| + | Normal procedure for PCR on both fragments all reactions where done as duplicates: | ||
| + | *Nir1 HF buffer | ||
| + | *Nir1 GC buffer | ||
| + | *Nir2 HF buffer | ||
| + | *Nir2 GC buffer | ||
| + | |||
| + | Both Nir1 reactions where done with primer pair 41 on 68C annealing and 2:00 min extension. | ||
| + | Both Nir2 reactions where done with primer pair 42 on 65C annealing and 2:30 min extension. | ||
| + | |||
| + | |||
| + | ===PCR on Nir1 with USER-primers=== | ||
| + | Samples where done i duplicates and with both HF- and GC-buffer. Nir1 as template and primer pair 39. 10 rounds of 60C annealing followed up by 25 rounds of 66C both had 5:00 min extension time. | ||
==Results== | ==Results== | ||
Revision as of 15:20, 4 August 2013
4 August 2013
Contents |
lab 208
Main purpose
Amplify the two Nir fragments and prepare for PCR with USER primers.
Who was in the lab
Kristian
Procedure
PCR for amplification of Nir fragments
Normal procedure for PCR on both fragments all reactions where done as duplicates:
- Nir1 HF buffer
- Nir1 GC buffer
- Nir2 HF buffer
- Nir2 GC buffer
Both Nir1 reactions where done with primer pair 41 on 68C annealing and 2:00 min extension. Both Nir2 reactions where done with primer pair 42 on 65C annealing and 2:30 min extension.
PCR on Nir1 with USER-primers
Samples where done i duplicates and with both HF- and GC-buffer. Nir1 as template and primer pair 39. 10 rounds of 60C annealing followed up by 25 rounds of 66C both had 5:00 min extension time.
Results
Conclusion
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