Team:Freiburg/Notebook/24 August

From 2011.igem.org

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Annealing 60-70
Annealing 60-70
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To confirm the PCR-Product has the correct size, load 2 µl of the sample onto an agarose-gel.
 
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How did you label the PCR-Product, where is it stored and what do you do next?
 
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{{:Team:Freiburg/Templates/footer}}
{{:Team:Freiburg/Templates/footer}}

Revision as of 21:12, 21 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

Digest of CcaR

Investigators: Jakob

  • 10µl BSA
  • 10µl NEB
  • 2µl DpnI
  • 38µl DNA

Digest for 2h at 37°C

Transformation of CcaR

Investigators: Jakob

Testdigest of quickchanged CcaS

Investigators: Julia

today I prepared 9 minipreps of the the quickchanged CcaS.
They were digested with EcoRI and loaded on a gel.
Those which are not cut twice were changed during PCR correctly.
We will send them for sequencing today.

Freiburg 8 25 2011 qS50 S51.Jpg
gel loaded with: Ladder 1 kb, qS50 a,b,c,d , qS51 a, b, c, d
lower lane:Ladder 1 kb qS51 e

positive clones are qS50b and qS51a.
concentration of minpreps: qS50b: 88,6 ng/μl and qS51a: 81,1 ng/μl.

blue light receptor

Transformation

Investigators: Sandra

Repeat of the transformation from yesterday with the Lovtap-NotGate-pSB1A3 vector.

PCR

Name: Sophie


Date: 24.08.11
Continue from Experiment: primer design (Date) 19.08.11

(Name) Sandra

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 P24
2.5µl Primer dw P79
1µl dNTPs of Template DNA
1µl DNA-Template M45
0.5 µl Phusion (add in the end)

What program do you use?

20 cycles with an annealing temperature of 65/68/71 ºC (gradient-PCR) and touchdown then 10 cycles with an annealing temperature of 65/70/75 ºC and touchdown
digested with DpnI


To confirm the PCR-Product has the correct size, load 2 µl of the sample onto an agarose-gel. Freiburg blue 24.08..jpg

How did you label the PCR-Product, where is it stored and what do you do next?
the PCR-products are labelled Not, nOt and noT and are stored in the "unverdaute Minipreps II"-box (or in the Gibson box?) If Sequencing is correct, I will do a new 3A assembly with the LovTAP-PCR product and an Amp-vector next.

red light receptor

NAME OF YOUR EXPERIMENT

Investigators:NAME



Lysis cassette

NAME OF YOUR EXPERIMENT

Investigators:NAME


Precipitator

NAME OF YOUR EXPERIMENT

PCR

Name:Rüdiger


Date: 24.08
Continue from Experiment (Date)-

(Name)

Project Name: Precipitator

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 P68Primer
2.5µl Primer dw P70Primer
1µl dNTPs of Template DNAPrimer
1µl DNA-Template GST-6-P-1 a,b,c,d,ePrimer
0.5 µl Phusion (add in the end)

What program do you use?

Annealing 60-70

                       

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