Team:Heidelberg/Templates/Del week11 LP

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Contents

11-07-2013

Amplification from FS_13s to FS_15s; 6.4 kb

PCR for amplification of DelLP, DelFG and DelEG; lane1=Ladder log2, lane2=delLP(11.07), lane3=DelFG(11.07), lane4=DelEG (Phusion flash, 11.07), lane5=DelEG (Phusion II,11.07), lane6=1kb ladder plus; DelLP=6.4kbp, DelFG=5.3kbp, DelEG=16.4kbp; run at 100 V, 0.8 % gel (TAE)
Gel after excision
Reaction of DelLP
what µl
D. acidovorans DSM-39 1
FS_13_short: (1/10) 1
FS_15_short: (1/10) 1
Phusion flash Master Mix 10
dd H2O 6
DMSO 1
Conditions of Del LP
Biorad MyCycler
Cycles temperature [°C] Time [s]
1 98 10
12 98 1
68 ↓ 0.5 5
72 2:10
18 98 1
66 5
72 2:10
1 72 10min
1 12 inf

Results:

  • There was a very weak band visible on the right height.
  • The band was cut out anyway, as we wanted to use it as template for a Re-PCR
  • The DNA was purified using QIAquick Gel Extration Kit.

12-07-2013

Re-PCR of DelLP (FS_13s to FS_15s; 6.4 kb; 11-07-2013)

PCR for amplification of DelFG (12.07;FS6+FS11) and re-amplification of DelLP(12.07;FS13short+FS15short); lane1=Ladder log2, lane2=DelFG(12.07;FS6+FS11long), lane3=DelFG(12.07;FS6+FS11short), lane4=DelLP, lane5=1kb ladder plus; DelLP=6.4kbp, DelEG=11.6 kbp; run at 100 V, 0.8 % gel (TAE)
Reaction of DelLP
what µl
Gel extracted fragment LP (11-07-2013) 2
FS_13_short: (1/10) 1
FS_15_long: (1/10) 1
Phusion flash Master Mix 10
dd H2O 5
DMSO 1
Conditions of Del LP
Biorad MyCycler
Cycles temperature [°C] Time [s]
1 98 10
12 98 1
68 ↓ 0.5 5
72 2:10
18 98 1
66 5
72 2:10
1 72 10min
1 12 inf

Results:

  • The Re-PCR did not work, no bands were visible on the gel.
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