Team:Queens-Canada/11 August 2010
From 2010.igem.org
August 11, 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.