Team:Heidelberg/Templates/Del week11 pSB4K5

From 2013.igem.org

08-07-2013

Amplification from FS_01 to FS_16; 4.2 kb

PCR for amplification of del fragments DelAE (08.07), pSB4K5 (08.07) and DelG (07.07); lane1=Marker, lanes2-4=DelAE (5.3kbp), lanes5-10=pSB4K5 (4.2kbp), lanes11-13:DelG (6.4kbp); run at 100 V, 0.8 % gel (TAE)
PCR for amplification of del fragments DelAE (08.07), pSB4K5 (08.07) and DelG (07.07), cut
Reaction

(2x50µl)

what µl
Template pSB4K5 1
FS_01: (1/10) 2.5
FS_16: (1/10) 2.5
Phusion Master Mix 25
DMSO 2.5
dd H2O 16.5


Conditions
Biorad MyCycler
Cycles-PCR temperature [°C] Time [s]
1 98 5
14 98 1
62 ↓ 0.5 5
72 3:20 min
16 98 1
60 5
72 3:20 min
1 72 12 min
1 4 inf

Results:

  • Amplification worked very well, we had bright bands on the right height.
  • Bands were cut out and DNA purified using QIAquick Gel Extration Kit.
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