Team:Heidelberg/Templates/Del week11 AG

From 2013.igem.org

12-07-2013

Amplification from FS_02 to FS_07; 16.4 kb

PCR for amplification of DelAG (12.07;FS02+FS07); lane6=Ladder log2, lane7=Ladder 1kb base ruler, lane8=DelAG(12.07;FS02+FS_07) run at 100 V, 0.8 % gel (TAE)
Reaction
what µl
D. acidovorans DSM-39 1
FS_02: (1/10) 2
FS_07: (1/10) 2
Phusion flash Master Mix 10
DMSO 1
dd H2O 4
Conditions
Biorad MyCycler
Cycles temperature [°C] Time [s]
1 98 10
12 98 1
68 ↓ 0.5 5
72 5:30
18 98 1
66 5
72 5:30
1 72 15min
1 12 inf

Results:

  • only a smear occured, no specific product was amplified
  • PCR will be repeated with a lower, constant annealing temperature