Team:Queens-Canada/27 July 2010

PCR

Hao and Nelson

pSip-1::1A3

Program run using 9:1 FAST:HIFI blend with an extension time of 6 s and annealing temperature of 55 °C.

Gel Electrophoresis

Thai and Yuli

pSB1A3, pSB1C3, pSB1T3, pSB1A2

Used 0.7 g agarose in 100 mL TBE. Gel run for 55 mins at 100 V.

Digestion

Hao and Nelson

pRab-7, unc-54 3'UTR, pStr-1, BBa_I13522, 1A2

Cut with XbaI and PstI. Treated with 1 µL CIAP. Only ~5µL of pRab-7, and ~9 µL of unc-54 3'UTR added in digestion. Both required 10 µL but ran out.

PCR

Yuli and Steve

eGFP

Program run using 9:1 FAST:HIFI blend with an extension time of 15 s and annealing temperature of 63 °C.

PCR

Yuli

pSip-1::eGFP, pRab-7::eGFP, pMec-7::eGFP

Re-ran pSip-1::eGFP, pMec-7::eGFP and pRab-7::eGFP from 22/07/10 for 10 cycles. Program run using 9:1 FAST:HIFI blend with an extension time of 45 s and annealing temperature of 61 °C.

Ligation

Hao

pRab-7, pStr-1, unc-54 3'UTR, pSB1A2, BBa_I13522

Heat Shock

Hao and Mary

pRab-7::1A2, pStr-1::1A2, unc-54 3'UTR::1A2, BBa_I13522

Transformed onto Amp plates.