Team:Queens-Canada/30 August 2010

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August 30, 2010

Liquid Culture

Thai

pOsm-10::eCFP::unc-54 3'UTR::1A3, pSra-10::eCFP::unc-54 3'UTR::1A3

Digestion

Hao and Thai

pSip-1::1C3, pFlp-1::1C3, pOdr-10::1C3, mCherry::1C3

EcoRI and SpeI XbaI and PstI EcoRI and PstI
pStr-220::1C3 eCFP::unc-54 3'UTR::1T3 pSB1T3
mCherry::1C3 unc-54 3'UTR::1C3
pFlp-1::1C3
pSip-1::1C3
pOdr-10::1C3

Backbone treated with CIAP for 30 mins.

Ligation

Hao and Thai

pSip-1, pStr-220, pFlp-1, pOdr-10::eCFP::unc-54 3'UTR::1A3, ChR2::unc-54 3'UTR::1T3

EcoRI and SpeI XbaI and PstI EcoRI and PstI
pStr-1 eCFP::unc-54 3'UTR 1A3
pStr-220 eCFP::unc-54 3'UTR 1A3
pFlp-1 eCFP::unc-54 3'UTR 1A3
pOdr-10 eCFP::unc-54 3'UTR 1A3
mCherry unc-54 3'UTR 1T3

Backbone treated with CIAP for 30 mins.

Heat Shock

Hao

pSip-1, pStr-220, pFlp-1, pOdr-10::eCFP::unc-54 3'UTR::1A3, ChR2::unc-54 3'UTR::1T3

Transformed ligation products from 30/08/10

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