Team:Queens-Canada/11 August 2010

PCR

Thai and Chris

ChR2::eYFP

Program run using 9:1 FAST:HIFI blend with an extension time of 20 s and annealing temperature of 65 °C.

Digestion-Ligation

Steve and Hao

pRab-7, pSip-1, pSB1C3, pFlp-1, pOdr-10, pSrb-6, pCeh-12, pSra-10, eYFP, ChR2, eCFP

pSB1C3 cut with XbaI and PstI, and EcoRI and PstI. pRab-7 and pSip-1 cut with XbaI and PstI. All other sequences cut with EcoRI and PstI. All products ligated into 1C3 using T4 ligase.

Gel Electrophoresis

Yuli

ChR2::eYFP, NpHR::eCFP, eGFP

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

Miniprep

Hao and Steve

pCeh-12::1C3, eCFP::1C3, pStr-1::1C3, pStr-220::1C3, pOsm-10::1C3, pOdr-1:1C3, NpHR::1C3

Digestion

Hao and Steve

pCeh-12::1C3, eCFP::1C3, pStr-1::1C3, pStr-220::1C3, pOsm-10::1C3, pOdr-1:1C3, NpHR::1C3

Cut with EcoRI and PstI.

Gel Electrophoresis

Hao and Steve

Used 0.7 g agarose in 100 mL TBE. Gel run for 40 mins at 100 V.

Gel Extraction

Yuli

ChR2::eYFP, eGFP

Extracted from gel run on 11/08/10.

Heat Shock

Chris Diagnostic Plasmid

Gel Electrophoresis

Mary and Louis and Hao

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

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

Gel Electrophoresis

Hao

pOdr-1::1C3, NpHR::1C3, ChR2::YFP, eGFP

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