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Team:Freiburg/Notebook/27 July
From 2011.igem.org
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(→PCR of Lov-tap and Not-gate) |
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What program do you use? | What program do you use? | ||
| - | 57°C auf 70°C ( | + | "57°C auf 70°C" (first annealing temperature: 58°C and then after 10 cycles 68°C) |
Revision as of 13:21, 27 July 2011
Contents |
Commons
Gel
Investigators: Sandra, Sophie
We loaded the PCR products of the vectors on a gel and measured the DNA concentration. DNA concentration were high and we diluted the vectors to get an endconcentration of 25ng/microl.
green light receptor
NAME OF YOUR EXPERIMENT
Investigators:NAME
blue light receptor
PCR of Lov-tap and Not-gate
Investigators: Sandra, Sophie
We repeated the PCR again, with higher temperatures this time. After PCR we digested with DpnI 37°C for 1 hour and then purified the DNA with PCR purifictaion kit. Afterwards we measured the DNA concentration and loaded 2 microl to a gel. PCR
| Name:Sandra, Sophie
| Date: 27.07.11 |
| Continue from Experiment: NOT-Gate and LovTAP with Gibson-overhangs (now we use a little bit higher temperature for primer annealing) (Date): 25.07.11
(Name): Sandra, Sophie | |
| Project Name: Blue Light | |
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 | NOT_G_up
LOV_G_up |
| 2.5µl | Primer dw | NOT_G_dw
LOV_G_dw |
| 1µl | dNTPs | of Template DNA |
| 1µl | DNA-Template | S35 (BBa_K322999)
S45 (Bba_Q04400) |
| 0.5 µl | Phusion (add in the end) |
What program do you use?
"57°C auf 70°C" (first annealing temperature: 58°C and then after 10 cycles 68°C)
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?
red light receptor
NAME OF YOUR EXPERIMENT
Investigators:NAME
Lysis cassette
NAME OF YOUR EXPERIMENT
Investigators:NAME
Precipitator
NAME OF YOUR EXPERIMENT
Investigators: NAME
