Team:Queens-Canada/10 August 2010

Gel Electrophoresis

Yuli

eGFP, pFlp-1

Used 0.4 g agarose in 60 mL TBE. Gel run for 45 mins at 100 V.

PCR

Hao

eYFP, mCherry, ChR2

Program run using 9:1 FAST:HIFI blend with an extension time of 15 s and annealing temperature of 65 °C.

eYFP and mCherry were run for 10 cycles without primers, then 20 cycles after primers were added. The Annealing temperature was changed to 61 °C once primers were added.

eCFP was run for 20 cycles.

Gel Electrophoresis

Hao

eYFP, mCherry, ChR2

Used 0.3 g agarose in 40 mL TBE. Gel run for 45 mins at 100 V.

Liquid Culture

Thai

pOsm-10, pStr-220, pOdr-1, NpHR, pStr-1, pCeh-12, eCFP

Topped up culture tues from 08/08/10 with 5 mL 2XTY + Chlor.

Heat Shock

Thai

pOsm-10, pStr-220, pOdr-1, NpHR, pStr-1, pCeh-12, eCFP

Heat Shock products placed onto C plates.

Gel Extraction

Hao

eYFP, mCherry, ChR2

Extraction performed on gel from 10/08/10. Used 400 µL binding buffer.

PCR

Yuli

ChR2, eYFP

Program run using 9:1 FAST:HIFI blend with an extension time of 20 s and annealing temperature of 62 °C.

PCR

Chris

NpHR::eCFP

Program run using 9:1 FAST:HIFI blend with an extension time of 25 s and annealing temperature of 65 °C.