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Team:Freiburg/Notebook/14 July
From 2011.igem.org
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Revision as of 14:57, 18 August 2011
Contact 
This is the wiki page of the Freiburger student team competing for iGEM 2011. Thank you for your interest!
Contents |
red light receptor
3A-assembly: Digest
Investigators: Jakob
3A-assembly of S22 (ho1)+S21a (terminator) and S23 (pcyA)+S21a (terminator)
S21a:
- Enzyme 1: XbaI
- Enzyme 2: PstI
S22, S23
- Enzyme 1: EcoRI
- Enzyme 2: SpeI
Vector:
- Enzyme 1: EcoRI
- Enzyme 2: PstI
- Enzyme 3: DpnI
3A-assembly: Ligation and transformation
Investigators: Julia
Ligation of S22 (ho1)+S21a (terminator) and S23 (pcyA)+S21a (terminator) in vector pSB1T3.
L23: Ligation of S23+S21a in pSB1T3 L22: Ligation of S22+S21a in pSB1T3
Plasmids were transformed in cells and plated out.
PCR
Investigators: Julia
PCR of the red light receptor, cph8.
Primer:
- P 10
- P 11
DNA template:
- BBa_I150010
Lysis cassette
Quickchange PCR
| Name: Theo
| Date: 14.07.2011 |
| Continue from Experiment Lysis Cassette
| |
| Project Name: Correct number of nucleotides between RBS and ATG of temp. sensitive repressor from
Lysis Device | |
PCR-Mixture for one Reaction:
For a 50 µl reaction use
| 32,5µl | H20 | Name |
| 10µl | 6x HF Phusion Buffer | |
| 2.5µl | Primer fw | P29 (1:10) |
| 2.5µl | Primer dw | P30 (1:10) |
| 1µl | dNTPs | |
| 1µl | DNA-Template | M15a (from S15->diluted to 5ng/µl) |
| 0.5 µl | Phusion (add in the end) |
What program do you use?
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Precipitator
Test-digest
Investigators: Rüdiger
Testdigest of the minipreps, Sophie did (june, 13).
- Enzyme 1: PstI
- Enzyme 2: EcoRI
Image of different samples from miniprep.
