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From 2011.igem.org

Contents 1 Commons 1.1 Gel 2 green light receptor 2.1 NAME OF YOUR EXPERIMENT 3 blue light receptor 3.1 PCR of Lov-tap and Not-gate 4 red light receptor 4.1 NAME OF YOUR EXPERIMENT 5 Lysis cassette 5.1 NAME OF YOUR EXPERIMENT 6 Precipitator 6.1 NAME OF YOUR EXPERIMENT

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