User:Jose Fabricio Lopez/Notebook/Logbook/2011/09/04: Difference between revisions

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|--
|--
|DNA diluited 10ng/μL
|DNA diluited 10ng/μL
|
|1μL
|1μL
|--
|Phusion
|.5μL
|.5μL
|--
|OligoFwd
|5μL
|5μL
|--
|OligoRev
|5μL
|5μL
|--
|H<sub>2</sub>O
|24.5μL
|24.5μL
|}
|}


Revision as of 15:54, 5 September 2011

iGEM 2011 <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page
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Goal

  • PCR for trouble shooting about PCRs for isothermal assembly.

Troubleshooting

  • PCR with Taq Platinum invitrogen and 57°C for annealing.

The objectives of those changes are:

  • Change times for elongation.
  • Change Temperature for anneling.
  • Excellent yield by PCR.

Gel

Completely unsuccessful

We are trying to know what is happening with PCRs.


PCR with different parameters

So far, we have amplified HydG and PFOR1, since now we are trying a new approach. For that we made a simply PCR, this is without GC buffer and DMSO. PCR is for HydEF1 and HydEF2

55°C 60°C
HydEF1 HydEF2
HydEF1 HydEF2
Reagents 55°C 60°C
Buffer HF Phusion 10μL 10μL
dNTPs (10mM each) 4μL 4μL
DNA diluited 10ng/μL 1μL 1μL
Phusion .5μL .5μL
OligoFwd 5μL 5μL
OligoRev 5μL 5μL
H2O 24.5μL 24.5μL



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