Team:Queens-Canada/28 July 2010

PCR

Hao

pSip-1, pRab-7

Program run using 9:1 FAST:HIFI blend with an extension time of 6 s and annealing temperature of 58 °C.

Gel Electrophoresis

Yuli and Steve

Used 1 g agarose in 140 mL TBE. Gel run for 35 mins at 100 V.

Digestion

Hao

pSB1A3

Half the samples were cut with EcoRI & SpeI, and the other half were cut with XbaI and PstI.

Gel Electrophoresis

Hao and Steve

pSB1A3

Used 0.5 g agarose in 70 mL TBE. Gel run for 40 mins at 100 V.

Digestion-Ligation

Steve and Thai

pRab-7, pSip-1, pMec-7, pSB1A3

Cut with XbaI and PstI. Backbone treated with CIAP.

Digestion

Hao and Nelson

pSrb-6, pCeh-12, pOdr-10, NpHR, ChR2, pOsm-10, pStr-1, eCFP, pHsp-3, pFlp-1, pSB1A3, pStr-220

Cut with EcoRI and PstI. May have damaged spin column filter of pHsp-3 during PCR purification. Transferred flow-through to new spin column. Old spin column may have trapped the DNA. Finished PCR purification with both columns.