Team:Heidelberg/Templates/Del week10 EG

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Revision as of 02:59, 30 September 2013 by Charly (Talk | contribs)

06-07-2013

Amplification from FS_04 to FS_07; 11.1 kb

PCR for amplification of del fragments E-G and A-E, AE+=PCR AE with DMSO, AE-=PCR without DMSO, M= Marker; wanted amplicon sizes are: AE=5.3kbp, EG=11.0


PCR for amplification of del fragments AE and EG, cut


Reaction


what µl
D. acidovorans DSM-39 1
FS_04: (1/10) 1
FS_07: (1/10) 1
Phusion Master Mix 10
DMSO 1/-
dd H2O 6/7



Conditions


Biorad MyCycler
Cycles-PCR temperature [°C] Time [s]
1 98 10
12 98 1
68 ↓ 0.5 5
72 3 min
18 98 1
66 5
72 3 min
1 72 10 min
1 4 inf


Results:

  • Amplification of DelEG worked but another unexpected product was amplified as well
  • band was cut out avoiding contamination with other product and DNA purified using QIAquick Gel Extraction Kit



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