Kun:LabNotes/ASE/2007-10-15: Difference between revisions

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(New page: ===Exp1: Fixing the PGP1Ver2 PCR reactions I did on 2007-10-12=== *Since I haven't run the thermalcycling on the 96 reactions on cDNAs, I'm just adding the ThermoStart Taq enzyme and more ...)
 
 
Line 6: Line 6:
     10mM dNTP        0.5    x100    50
     10mM dNTP        0.5    x100    50
     H2O              0.15    x100    15
     H2O              0.15    x100    15
<b> I used the aluminum tape to seal the plate. It turned out that I had a evaporation issue. I don't think this plate is useful.</b>


*Also set up 32 reactions on the genomic DNA template.  
*Also set up 32 reactions on the genomic DNA template.  
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95C 15min -> 40 cycles of (94C 30S -> 56C 30S -> 72C 1min) -> 72C 3min, using the PTC-200 machine.

Latest revision as of 18:19, 16 October 2007

Exp1: Fixing the PGP1Ver2 PCR reactions I did on 2007-10-12[edit]

  • Since I haven't run the thermalcycling on the 96 reactions on cDNAs, I'm just adding the ThermoStart Taq enzyme and more ABI's dNTPs into the reactions.
   10x buffer       0.1     x100     10
   ThermoStart Taq  0.25    x100     25
   10mM dNTP        0.5     x100     50
   H2O              0.15    x100     15

I used the aluminum tape to seal the plate. It turned out that I had a evaporation issue. I don't think this plate is useful.

  • Also set up 32 reactions on the genomic DNA template.

Reaction set up:

' one reaction 33 reactions
Template 1 each
10x Buffer 5 165
25mM MgCl2 3 100
10mM dNTP 1 33
10uM primer mix 1 each
50x SYBG I 0.4 13.2
ThermoStart Taq 0.25 8.25
H2O 38.35 1267

95C 15min -> 40 cycles of (94C 30S -> 56C 30S -> 72C 1min) -> 72C 3min, using the PTC-200 machine.