Team:Paris Saclay/Notebook/September/16

From 2014.igem.org

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[https://2014.igem.org/Team:Paris_Saclay/Notebook/September/15#PCR see here]
[https://2014.igem.org/Team:Paris_Saclay/Notebook/September/15#PCR see here]
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[[File:1609 chromo topo 2-9 jaune 10-17 bleu.jpg|400px]
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[[File:1609 chromo topo 2-9 jaune 10-17 bleu.jpg|400px]]
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well 2-9 = chromo yellow in topo plasmid
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well 10-17 = chromo blue in topo plasmid

Revision as of 12:05, 26 September 2014

Contents

Lab Work

By Mélanie

D- Lemon scent

Plasmids extraction

Extraction of the plasmid from the culture made yesterday(PS)

Digestion of PS topo plasmids

by PacI and PSTI

1609 digestion ps et gs par pacI.jpg

(well 2 and 3 : PacI digestion)

Ligation

After digestion, ligation overnight in PPS1


PCR Colony

Test of the transformation made yesterday to know if we had obtained a plasmid]

1609 PCR sur clones.jpg

primer to ckeck the direction of the insert PPS2 : iPS72-96

PPS4 : ips72-97

= no insert (just well 1-9)

liquide culture

liquide culture from yesterday's colony to make stock tomorrow

PCR cad to cloning in pSBIC3

PCR and digestion PSTI/ecoRI and ligation

PCR GS

PCR to check if we have the insert in our topo plasmid

see here


B- Construction of the fusion protein (color)

PCR

PCR to check if we have the insert in our topo plasmid

see here

1609 chromo topo 2-9 jaune 10-17 bleu.jpg

well 2-9 = chromo yellow in topo plasmid well 10-17 = chromo blue in topo plasmid