Team:Heidelberg/Templates/Del week10 L

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Revision as of 01:05, 30 September 2013

Contents

04-07-2013

Amplification from FS_14 to FS_15; 1.4 kb

Gel eletrophoresis of PCRs of DelF-G, DelO-P and DelL


Reaction
what µl
D. acidovorans DSM-39 1
FS_14: (1/10) 1
FS_15: (1/10) 1
Phusion Master Mix 10
DMSO 1
dd H2O 6
Conditions
Biorad MyCycler
Cycles temperature [°C] Time [s]
1 98 10
30 98 1
72 40
1 72 7 min
1 4 inf

Results:

  • Amplification worked very well, we had a bright band on the right height.
  • Band was cut out and DNA purified using QIAquick Gel Extration Kit.

06-07-2013

Amplification from FS_14 to FS_15; 1.4 kb

PCR for amplification of del fragment L, L+=PCR delL with DMSO, L-=PCR without DMSO, M= Marker run at 135 V, 0.8 % gel (TAE); wanted amplicon size is: 1.4 kbp
Reaction
what µl
D. acidovorans DSM-39 1
FS_14: (1/10) 1
FS_15: (1/10) 1
Phusion Master Mix 10
DMSO 1/-
dd H2O 6/7
PCR for amplification of del fragment L, cut
Conditions
Biorad MyCycler
Cycles-PCR temperature [°C] Time [s]
1 98 10
12 98 1
66 ↓ 0.5 5
72 42
18 98 1
63 5
72 42
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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