Team:Heidelberg/Templates/Del week15 pSB4K5
From 2013.igem.org
Contents |
07-08-2013
Results Sequencing
Send the backbone pSB4K5 from the partsregistry to GATC for sequencing:
- pSB4K5 with Primer FS_01
- pSB4K5 with Primer FS_16
Results:
- The sequence is the same as expected and indicated in the partsregistry.
Amplification from FS_01 to FS_16; 4.2 kb
- 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 T100 | ||
---|---|---|
Cycles-PCR | temperature [°C] | Time [s] |
1 | 98 | 5 |
12 | 98 | 1 |
62 ↓ 0.5 | 5 | |
72 | 1:30 min | |
18 | 98 | 1 |
60 | 5 | |
72 | 1:30 min | |
1 | 72 | 5 min |
1 | 8 | 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.
08-08-2013
Concentration measurement
The concentration of the gel purified fragment was measured using a NanoDrop Instrument.
Fragment | Primer | Date PCR | Concentration |
---|---|---|---|
pSB4K5 | FS16-FS01 | 07-08-2013 | 55.9 ng/µl |