Team:Freiburg/Notebook/6 September

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Revision as of 19:12, 6 September 2011


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

Contents

Commons

Miniprep

Investigators: Sandra

Yesterday LB was inoculated with strains (S14, S39-S44 etc.). Miniprep of strains.


Digestion: 2A-assembly

Investigators: Sandra

Digestion of:

  • S14 (GFP)

with XbaI and PstI

  • S39 (PR1)
  • S40 (PR2)
  • S41 (PR3)
  • S42 (PR4)
  • S43 (PR5)
  • S44 (PR6)

with SpeI and PstI

incubation for 6 hours at 37°C.

green light receptor

NAME OF YOUR EXPERIMENT

Investigators:NAME


blue light receptor

Trafo

Investigators: Sandra

Trafo of ligated parts of the 2A-assembly.

Sequence analysis

sequence analysis of our lovtap-Notgate was neagtive:-( So we´ll try the gibson assembly.

PCR

Investigators: Sophie PCR


Name: Sophie


Date: 06.09.11
Continue from Experiment (Date)

(Name)

Project Name: Gibson of ♥ and NOT

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 ♥: P97

NOT: P99

2.5µl Primer dw ♥: P98

NOT: P100

1µl dNTPs of Template DNA
1µl DNA-Template NoT, M45 (BBa_Q0400)

original DNA from Edinburgh (BBa_K322999)

0.5 µl Phusion (add in the end)

Digestion with Dpn I and purification with PCR purification kit What program do you use?

First 20 cycles touchdown 69°C -0.4/cycle

next 10 cycles touchdown 72°C -0.2 per cycle


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?

red light receptor

NAME OF YOUR EXPERIMENT

Investigators:NAME



Lysis cassette

Troubleshooting of the modified Lysis genes K124017

Investigators:Theo


Minipreps of the first inoculations were applied on a 1% agarose gel without digesting them to see if they have an insert of the modified Lysis genes K124017. Nr1, 5 and 7 were sent for sequencing since the size seems to be about right.

  • Note: S4 is the B0034 RBS (ribosome binding site), S15 is the non-modified K124017 Lysis Cassette without RBS


Freiburg11 9 8 2011 S4und15Verdau M66Verdau.Jpg


Precipitator

NAME OF YOUR EXPERIMENT

Investigators: NAME

                       

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