Team:Freiburg/Notebook/24 August

From 2011.igem.org

(Difference between revisions)
(→Precipitator)
 
(One intermediate revision not shown)
Line 1: Line 1:
{{:Team:Freiburg/Templates/header}}
{{:Team:Freiburg/Templates/header}}
 +
<html>
 +
<div id="notebook-page-header">
 +
<div id="notebook-back" width="100px" >
 +
<a href="https://2011.igem.org/Team:Freiburg/Notebook/23_August">Previous entry</a>
 +
</div>
 +
<div id="notebook-title">
 +
<a href="https://2011.igem.org/Team:Freiburg/Notebook"> 24 August </a>
 +
</div>
 +
<div id="notebook-next">
 +
<a href="https://2011.igem.org/Team:Freiburg/Notebook/25_August">Next entry</a>
 +
</div>
 +
</div>
 +
</html>
 +
 +
==<span style="color:green;">green light receptor</span>==
==<span style="color:green;">green light receptor</span>==
Line 199: Line 214:
Annealing 60-70
Annealing 60-70
-
 
-
 
-
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?
 
-
 
-
 
{{:Team:Freiburg/Templates/footer}}
{{:Team:Freiburg/Templates/footer}}

Latest revision as of 01:11, 22 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

                       

Retrieved from "http://2011.igem.org/Team:Freiburg/Notebook/24_August"