Team:Freiburg/Notebook/15 August

From 2011.igem.org

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(→NAME OF YOUR EXPERIMENT: Transformation)
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Project name: inducible promoter for pbd<br/>
Project name: inducible promoter for pbd<br/>
samples stored in incubator
samples stored in incubator
 +
 +
===PCR===
 +
 +
'''Investigators: Rüdiger'''
 +
 +
PCR of precipitator.
 +
 +
Primer:
 +
*p44
 +
*p45
 +
*p46
 +
*p48
 +
*51
 +
*52
 +
*53
 +
 +
Primer:
 +
*p54
 +
*p47
 +
 +
{| cellpadding="10" cellspacing="0" border="1"
 +
|name
 +
|primer
 +
|primer
 +
|-
 +
|44
 +
|p44
 +
|p54
 +
|-
 +
|45
 +
|p45
 +
|p54
 +
|-
 +
|46
 +
|p46
 +
|p54
 +
|-
 +
|48
 +
|p48
 +
|p47
 +
|-
 +
|51
 +
|p51
 +
|p54
 +
|-
 +
|52
 +
|p52
 +
|p54
 +
|-
 +
|53
 +
|p53
 +
|p54
 +
|}
{{:Team:Freiburg/Templates/footer}}
{{:Team:Freiburg/Templates/footer}}

Revision as of 13:17, 13 September 2011


This is the wiki page
of the Freiburger student
team competing for iGEM 2011.
Thank you for your interest!

Contents

green light receptor

Order primers

Investigators: Jakob

  • Order new primers for the quickchange of CcaS and CcaR

blue light receptor

Transformation

Investigators: Sophie

Transformation of ligated parts:

  • LovTAP
  • Not-Gate
  • T3 vector


stored in incubator at 37°C.

red light receptor

PCR for Cph8

Investigators:Julia


after protocol from Uppsala. PCR-Mixture for one Reaction:

For a 50 µl reaction use


32,5µl H20
10µl 5x Phusion Buffer
2.5µl Primer fw
2.5µl Primer dw
1µl dNTPs
1µl DNA-Template
0.5 µl Phusion (add in the end)

Primer used:
cph8 Prefix: TTCGAATTCGCGGCCGCTTCTAGATGGCCACCACCGTACAA
cph8 Suffix: CCGCTACTAGTATTATTACCCTTCTTTTGTCATGCCCT

PCR program:
Temp. Time,Nr of cycles
98° 5' 1x
98° 30 10x
65° 30
72° 1'15
98° 30 20x
72° 1'15
72° 7' 1x
4° ∞

Template: Voights plasmid (10x dilution)
Amplicon: 2235 bp

Elongation time Phusion 67,05 sec

Lysis cassette

NAME OF YOUR EXPERIMENT

Investigators:NAME


Precipitator

Ligation

Investigators: Sophie
continue from experiment: Cloning (12.8.11)
Project name: inducible promoter for pbd
Vector: psb1T3
Inserts: IPTG-Promoter,Pbd-GFP
samples stored in Ruediger's box

Transformation

Investigators: Sophie
continue from experiment: Ligation
Project name: inducible promoter for pbd
samples stored in incubator

PCR

Investigators: Rüdiger

PCR of precipitator.

Primer:

  • p44
  • p45
  • p46
  • p48
  • 51
  • 52
  • 53

Primer:

  • p54
  • p47
name primer primer
44 p44 p54
45 p45 p54
46 p46 p54
48 p48 p47
51 p51 p54
52 p52 p54
53 p53 p54

                       

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