Team:Queens-Canada/12 August 2010

PCR

Hao and Thai

pGpd-2, pHsp-3

Program run using 9:1 FAST:HIFI blend with an extension time of 25 s and annealing temperature of 57 °C.

Heat Shock

Hao and Thai Digestion-Ligation products from 11/08/10, unc-54 3'UTR::1A3

Used Amp plates for unc-54 3'UTR::1A3 and Chlor plates for all others.

Digestion

Chris

pStr-220::1C3

Cut with EcoRI and PstI.

Digestion-Ligation

Yuli and Steve

ChR2, unc-54 3'UTR, pSB1T3, pSB1C3, ChR2::YFP, eGFP

unc-54 3'UTR cut with XbaI and PstI. ChR2::YFP cut with EcoRI and SpeI. All others cut with EcoRI and PstI. Digestion products ligated into 1T3 and 1C3.

Gel Electrophoresis

Hao

pStr-220, pHsp-3, pGpd-2, ChR2::eYFP::unc-54 3'UTR::1T3, ChR2::eYFP::1C3, 1T3, 1C3, eGFP::1C3

Used 0.7 g agarose in 100 mL TBE. Gel run for 60 mins at 100 V.

Heat Shock

Yuli and Steve

ChR2::eYFP::unc-54 3'UTR::1T3, ChR2::eYFP::unc-54 3'UTR::1T3, ChR2::eYFP::1C3, eGFP::1C3

Microinjection

Yuli

Prepared injection pads.