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Team:Groningen/Labwork/16 July 2013
From 2013.igem.org
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<br/>did a Restriction digestion for Silk and Signal Sequences. incubation started at 11:00 for the signal sequences and 11:30 for the silk. | <br/>did a Restriction digestion for Silk and Signal Sequences. incubation started at 11:00 for the signal sequences and 11:30 for the silk. | ||
<br/> at 15:38 the restriction digestion reaction was run over a 1,5 % agarose gel with 100 v for 22 min. | <br/> at 15:38 the restriction digestion reaction was run over a 1,5 % agarose gel with 100 v for 22 min. | ||
| + | <br/> no result on the gel for the sik. resulst as expected for signal sequence. | ||
Revision as of 15:41, 16 July 2013
Mirjam
Finally the colonies are growing on LB agar plates with ampicillin. The negative control does not show any colonies, the double negative does??, the positive control shows a lot of red colonies. On one of the four plates there are no colonies present, on two of them only a couple and on one a lot.
The colonies are picked and placed in LB medium with ampicillin to grow further for miniprep. Colony PCR revealed no bands.
Mini prep analysis is done with the colonies of plate 1 and 3.
Restriction analysis is done with EcoRV.
Inne
Did a PCR with 3 samples in duplo
| Sample | Primer F | Primer R | Annealing temp. |
| 1 | without strep tag | without strep tag | 78 C |
| 2 | with strep tag | without strep tag | 78 C |
| 3 | without strep tag | with strep tag | 80 C |
Protocol:
| amount | compound |
| 28.2 uL | MilliQ |
| 1.5 uL | 5% DMSO |
| 1 uL | each dNTP |
| 10 uL | Phusion GC buffer |
| 2.5 uL | Primer F |
| 2.5 uL | Primer R |
| 1 uL | Silk template BBa_16 |
| 0.3 uL | Phusion polymerase |
Ran the gel 1.5% agarose using the big slots,
gel ran ADD 90 V for 45 minutes.
gel didn't run nice, tried to cut out bands from 2 and 3.
did gel purification
Ran 0.8% agarose gel for 24 min on 100 V
made new KBE buffer. 100ml KBE/900ml demiwater.
Sander
did a Restriction digestion for Silk and Signal Sequences. incubation started at 11:00 for the signal sequences and 11:30 for the silk.
at 15:38 the restriction digestion reaction was run over a 1,5 % agarose gel with 100 v for 22 min.
no result on the gel for the sik. resulst as expected for signal sequence.
