Team:Queens-Canada/6 August 2010

Gel Electrophoresis

Hao

pSra-10, pHsp-3

Used 0.3 g agarose in 40 mL TBE. Gel run for 50 mins at 100 V.

PCR

Hao and Thai

ChR2, eCFP, eYFP, mCherry

Program run using 9:1 FAST:HIFI blend with an extension time of 15 s and annealing temperature of 61 °C.

mCherry was run for 5 cycles without primers, then for 35 cycles with primers.

Ligation

Hao and Steve and Thai

NpHR, pOdr-10, eYFP, pStr-220, mCherry, pOsm-10, pOdr-1, eCFP, pMec-7, pFlp-1, pCeh-12, pIC518, ChR2, pSrb-6, pSB1C3

All inserts ligated into pSB1C3. T4 ligase used.

Heat Shock

Hao and Thai

Ligation products from 06/08/10

Gel Electrophoresis

Hao and Thai

eYFP,ChR2, mCherry

Used 0.3 g agarose in 45 mL TBE. Gel run for 45 mins at 100 V.