User:Paulina Alatriste/Notebook/UNAM Genomics Mexico 2011/2011/09/20

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Parts assembly

ABSTRACT

  • PCR to amplify HydEF1

  • I did two PCR reactions in order to see which annealing temperature is best to amplify HydEF1. I also change some reagents such as the Buffer for the polymerase. I use the following proportions:
H2O 22.8μL
Buffer GC 10μL
dATP 1 μL
dGTP 1 μL
dTTP 1 μL
dCTP 1 μL
Primer Fwd 5 μL
Primer Rev 5 μL
DNA template 1.2 μL
DMSO 1.5 μL
Phusion Pol 0.5 μL


Cycles Gel Lanes Gel
  • 95°C 5min
  • 98°C 10s 30Cy
  • 60°C-62°C 30S 30Cy
  • 72°C 2 min 30Cy
  • 72°C 10min
  • 4°C
  • Lane1 Ladder
  • Lane2 HydEF1 60°C
  • Lane3 HydEF1 62°C


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