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Team:Freiburg/Notebook/19 July
From 2011.igem.org
Contents |
green light receptor
NAME OF YOUR EXPERIMENT
Investigators:NAME
blue light receptor
Theoretical Gibson-Assembly
Investigators: Sandra, Sophie
Primer Degin for Blue light sensor (lovTAP) + trp promotor (BBa_K322999) and tetR Gen + tetO promotor (BBa_Q04400).
- LOVtap-Trp_up: gaattcgcggccgcttctagtcacacaggaaagtactatgt
- LOVtap-Trp_dw: tacttttatctaatctggacatctagtatttctcctctttgtcgataccctttttacgtg
- TetR-TetO_up: aaagaggagaaatactagatgtccagattag
- TetR-TetO_dw^: ctgcagcggccgctactag
^we also have another primer for this one, but we are not sure about the length/melting temperature. Might be changed.
red light receptor
NAME OF YOUR EXPERIMENT
Investigators:NAME
Lysis cassette
NAME OF YOUR EXPERIMENT
Investigators:NAME
Precipitator
NAME OF YOUR EXPERIMENT
PCR
| Name: Ruediger
| Date: 19.07.11 |
| Continue from Experiment (Date)
PCR 18.07 (Name) | |
| Project Name: GFP Pbd | |
PCR-Mixture for one Reaction:
For a 50 µl reaction use
| 32,5µl | H20 | Name |
| 10µl | 5x Phusion Buffer | of Primer |
| 2.5µl | Primer fw | P18, P19, P20 |
| 2.5µl | Primer dw | P28 |
| 1µl | dNTPs | of Template DNA |
| 1µl | DNA-Template | PCR product of P1,P3,S14 from yesterday |
| 0.5 µl | Phusion (add in the end) |
What program do you use?
One set of probes was prepared and then split into 3 tubes each to test them at Annealingtemperatures 44C, 52C and 60C for the first 10 cycles. Then Annealingtemperature 60C for 25 cycles
To confirm the PCR-Product has the correct size, load 2 µl of the sample onto an agarose-gel.
How did you label the PCR-Product, where is it stored and what do you do next?
S14+P18+P28
S14+P19+P28
S14+P20+P28
Stored in PCR product box
Lane1
Quick Load Marker
Lane2
44C S14+P18+P28
Lane3
44C S14+P19+P28
Lane4
44C S14+P20+P28
Lane5
52C S14+P18+P28
Lane6
52C S14+P19+P28
Lane7
52C S14+P20+P28
Lane8
60 S14+P18+P28
Lane9
60 S14+P19+P28
Lane10
60 S14+P20+P28
