Team:Heidelberg/Templates/Del week16 G

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Revision as of 01:00, 30 September 2013

Contents

15-08-2013

Amplification from SR_01 to FS_23; 6.2 kb

Amplification of DelG; run at 100 V, 0.8 % gel (TAE)(14.08)
Amplification of DelG; run at 100 V, 0.8 % gel (TAE)(14.08)
Reaction (5x20 µl)
what µl
D. acidovorans SPH-1 colony 1
SR_01: (1/10) 2
FS_23: (1/10) 2
Phusion flash Master Mix 10
DMSO 1
dd H2O 4
Conditions
Biorad T100
Cycles temperature [°C] Time [s]
1 98 10
12 98 1
66 ↓ 0.5 5
72 3:20
18 98 1
64 5
72 3:20
1 72 10min
1 12 inf

Results:

  • Amplification of DelG was successful
  • Bands were cut out and DNA purified using QIAquick Gel Extraction Kit
  • Amplification will be repeated to increase the concentration for the Gibson Assembly

16-08-2013

Concentration measurement

the fragment was gel purified

Fragment Primer Date PCR Concentration
DelG SR01-FS23 15-08-2013 52.6 ng/µl
DelG after nucleotide removal SR01-FS23 15-08-2013 30 ng/µl

Amplification from SR_01 to FS_23; 6.2 kb

Amplification of DelG ; run at 100 V, 0.8 % gel (TAE)(14.08)
Amplification of DelG ; run at 100 V, 0.8 % gel (TAE)(14.08)
Reaction (5x20 µl)
what µl
D. acidovorans SPH-1 colony 1
SR_01: (1/10) 2
FS_23: (1/10) 2
Phusion flash Master Mix 10
DMSO 1
dd H2O 4
Conditions
Biorad T100
Cycles temperature [°C] Time [s]
1 98 10
12 98 1
66 ↓ 0.5 5
72 3:20
18 98 1
64 5
72 3:20
1 72 10min
1 12 inf

Results:

  • Amplification of DelG was successful
  • Bands were cut out and DNA purified using QIAquick Gel Extraction Kit
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