Team:Paris/Addressing testing

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iGEM > Paris > WetLab > Addressing

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WetLab

Main - Addressing - Production - reception - Addressing

Adressing system

global constructions :

Time required : A lOT !!!!!

Experiments ran :

row 1	row 2	row 3
PCR	PCR	PCR
[]	2003	Miller SI & Guina T.
[]	2005	Kuehn MJ & Kesty NC.
[]	2006	McBroom AJ & Kuehn MJ.
[]	2007	McBroom AJ & Kuehn MJ.
[]	2009	Deatherage BL & Cookson BT.
[]	2009	Purnick PE & Weiss R.

From plate: mRFP

PCR :

ClyA Cterm plasmid : ? Oligo : TM : ClyA Nterm plasmid : ? Oligo : TM : mRFP Cterm plasmid : ? Oligo : TM : mRFP Nterm plasmid : ? Oligo : TM :

Verification on gel

ok

Purification on gel

ok

digestion:

ClyA Cterm S/P RFP Nterm X/P PBAD E/S

ligation:

ClyA Cter-RFP Nter vector PSB1A3

PCR colo :

miniprep:

verifivation of the miniprep by digestion

sequencing [[ClyA]]

-ClyA Nterm= biobrick+sequencage+stock glycerol Done

-ClyA Cterm= biobrick+sequencage+stock glycerol Done

[[-RFP-]]

-RFP Nterm= biobrick+sequencage+stock glycerol Done

-RFP Cterm= biobrick+sequencage+stock glycerol Done

[[-ClyA Cter-RFP Nter-]]

-ClyA Cter-RFP Nter= biobrick+stock glycerol Done

[[-RFP Cter-ClyA Nter-]]

-miss ligation between 2 parts (j'ai fait plusieurs ligation le 7ou8/09, mais pas marché le 9/09 apres culture overnight, voir si les autres ont continué)

[[PBad-ClyA Cter-RFP Nter-]]

-failed many time with:

Pbad cut in S/P and ClyA Cter-RFP Nter in X/P

Pbad cut in E/S and ClyA Cter-RFP Nter in E/X

Export system

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 |2009
 |Purnick PE & Weiss R.
+|}
 From plate: mRFP