Team:Freiburg/Notebook/19 July

From 2011.igem.org

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(NAME OF YOUR EXPERIMENT)
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===NAME OF YOUR EXPERIMENT===
 
'''PCR'''
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Stored in PCR product box
Stored in PCR product box
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[[File:Freiburg11_7_20_2011_2_48_01_PM.Jpg]]
 
[[File:Freiburg11_Phusion_ladder.JPG]]
[[File:Freiburg11_Phusion_ladder.JPG]]
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[[File:Freiburg11_7_20_2011_2_48_01_PM.Jpg]]
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Lane1
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Quick Load Marker
Quick Load Marker

Revision as of 16:34, 19 July 2011

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

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

Freiburg11 Phusion ladder.JPG Freiburg11 7 20 2011 2 48 01 PM.Jpg

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