Team:Queens-Canada/31 August 2010

From 2010.igem.org

August 31, 2010

Miniprep

Hao and Thai

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

Digestion

Hao and Thai

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

Gel Electrophoresis

Hao and Thai

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

Used 0.7 g agarose in 100 mL TBE. Gel run for 30 mins at 100 V.

Digestion

Hao and Chris PRF4, eYFP, eGFP

Gel Electrophoresis

Hao and Chris PRF4

Used 0.3 g agarose in 40 mL TBE. Gel run for 40 mins at 100 V.

Ligation

Hao and Chris

eGFP::unc-54 3'UTR::1A3, eYFP::unc-54 3'UTR::1A3, ChR2::unc-54 3'UTR::1A3

EcoRI and SpeI XbaI and PstI EcoRI and PstI
eYFP unc-54 3'UTR 1A3
eGFP unc-54 3'UTR 1A3
mCherry unc-54 3'UTR 1A3

Backbone treated with CIAP for 30 mins.

Heat Shock

Hao and Chris

eGFP::unc-54 3'UTR::1A3, eYFP::unc-54 3'UTR::1A3, mCherry::unc-54 3'UTR::1A3