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Revision as of 16:34, 19 July 2011

Contents 1 green light receptor 1.1 NAME OF YOUR EXPERIMENT 2 blue light receptor 2.1 Theoretical Gibson-Assembly 3 red light receptor 3.1 NAME OF YOUR EXPERIMENT 4 Lysis cassette 4.1 NAME OF YOUR EXPERIMENT 5 Precipitator 5.1 NAME OF YOUR EXPERIMENT

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