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Team:Freiburg/Notebook/15 July
From 2011.igem.org
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Strange band size in lane 3. 4 and 5 did not work. | Strange band size in lane 3. 4 and 5 did not work. | ||
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Revision as of 15:03, 18 July 2011
Contents |
Meeting
attendants: Tobi, Theo, Julia, Rüdiger, Jakob, Sandra
green light receptor
already done:
To-do:
blue light receptor
already done:
- Transformation of Lov-Tap in cells.
To-do:
- Cloning of promotor (medium) in Lov-Tap part and perform a transformation. Plating out of the cells on trypthophan-free medium.
red light receptor
already done:
To-do:
Lysis cassette
Quick change
Investigators: Theo
already done:
- repressor part (BBa_K098995) has no bases between RBS (B0034) and start codon (K098997), resulting in no translation.
- quickchange of the repressor part to insert 6bp between RBS and start codon
To-do:
- DpnI digestion to digest the DNA template (methylated DNA) of the PCR to have only the new synthesized DNA strand.
- After digestion with DpnI, transformation of cells with the corrected part (BBa_K08995)
Precipitator
PCR
| Name:
Ruediger | Date:
18.07.2011 |
| Continue from Experiment (Date)PCR 1507
(Name) Ruediger | |
| Project Name:
GFP Pbd | |
PCR-Mixture for one Reaction:
For a 50 µl reaction use
| 32,5µl | H20 | |
| 10µl | 5x Phusion Buffer | |
| 2.5µl | Primer fw | |
| 2.5µl | Primer dw | |
| 1µl | dNTPs | |
| 1µl | DNA-Template | |
| 0.5 µl | Phusion (add in the end) |
What program do you use?
10x (95C-41/52C-72C) + 25x ((95C-60C-72C)
How did you label the PCR-Product, where is it stored and what do you do next?
Reactions:
P28+P18+M14.1
P28+P19+M14.1
P28+P20+M14.1
Results:
did not work well.
Strange band size in lane 3. 4 and 5 did not work.
