Team:Freiburg/Notebook/1 September

From 2011.igem.org

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'''Investigators:Theo'''
'''Investigators:Theo'''
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Another miniprep of the parts was done
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The digestion of the parts:
*S66 = modified Lysis genes K124017 in pSB1A2
*S66 = modified Lysis genes K124017 in pSB1A2
*GFP = E0040 in pSB1A2
*GFP = E0040 in pSB1A2
*S48 = Quickchange-modified K098995 in pSB1A2
*S48 = Quickchange-modified K098995 in pSB1A2
*S39 = Promotor (J23104) + RBS (B0034) in pSB1C3
*S39 = Promotor (J23104) + RBS (B0034) in pSB1C3
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was run on the gel to see if the inserts are there.
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As we thought, there is no band for S66, although there is DNA isolated from the mini-prep. It is probably a matter of making a stock from the false mini-prep. More answers on Monday or Tuesday
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There is no band for S66 so we sent it for sequencing
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[[File:9_6_2011_Minis_von_S66_GFP_S48_S39.Jpg‎]]
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[[File:9_2_2011_9_43_43_AM.Jpg‎‎]]
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Revision as of 15:09, 4 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

NAME OF YOUR EXPERIMENT

Investigators:NAME


blue light receptor

Picking clones

Investigators: Sandra

There were several clones on the cm plates. Clones were picked and incubated in LB cm medium at 37°C.

Gibson primer design

Investigators: Sophie, Sandra, Ruediger
we designed some new gibson primers for the assembly of NOT and LOV-tap in a vector because we are not sure if we can achieve this by 3A assemblies.

red light receptor

NAME OF YOUR EXPERIMENT

Investigators:NAME



Lysis cassette

3A Assembly of Quickchange-modified K098995 + modified Lysis genes K124017

Investigators:Theo
The digestion of the parts:

  • S66 = modified Lysis genes K124017 in pSB1A2
  • GFP = E0040 in pSB1A2
  • S48 = Quickchange-modified K098995 in pSB1A2
  • S39 = Promotor (J23104) + RBS (B0034) in pSB1C3

was run on the gel to see if the inserts are there.


There is no band for S66 so we sent it for sequencing



9 2 2011 9 43 43 AM.Jpg

Precipitator

Ligation

Name: Sophie Date: 1.9.11
Continue from Date: 31.08.11 Name: Sophie

Experiment: Digestion

Project Name:

Procedure


PCR tube:

total volume 20 μl


  1. add H2O (17 μl -X-Y-Z)
  2. add 2 μl Ligase Buffer 10x
  3. add Insert 1, Insert 2(when proceeding from 3A digestion use 2 μl of each)
  4. add Vector (20ng needed. When proceeding from 3A digestion use 2 μl)
  5. Add 1 μl T4-DNA Ligase
  6. Incubate 10-30 min at room temperature
  7. heat for 20 minutes at 80°C
  8. store at -20°C or directly proceed to transformation


Name of part Ratio Insert:Vector

= 3:1 or 1:1

Volume (μl)
X insert 1 GFP-pbd-PCR a/b both
Y vector pGEX
H2O

Documentation:

Why are you doing this experiment? Where are your parts stored? Name the parts for ligation etc.


Why? To get data about the plastic binding for modeling

stored in: “Minipreps, verdaut”-box

ligation-products stored in: ligation box

parts for ligation: GFP-pbd-PCR a/b, pGEX


                       

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