Team:Queens-Canada/4 August 2010

Gel Electrophoresis

Basia and Thai

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

Heat Shock

Hao and Basia

Re-performed heat shock from 03/08/10 with leftover TOP10 cells and additional 5 µL of ligation product. Prepared new LB plates with Chlor antibiotics. Before plating, tubes were spun down at 13000g for 2 mins, the supernatant was discarded, and pellets were resuspended in 200µL of 2XTY.

Miniprep

Hao and Thai

pStr-1::1A2, unc-54 3'UTR::1A3, pRab-7::1A3

HB Buffer was not used (not crucial).

PCR

Thai and Hao

pHsp-3, pSra-10

Program run using 9:1 FAST:HIFI blend with an extension time of 33 s and annealing temperature of 59 °C.

Digestion

Thai and Mary

Cut with EcoRI and PstI.

Gel Electrophoresis

Thai and Mary

pStr-1::1A2, unc-54 3'UTR::1A3, pRab-7::1A3

Used 0.7 g agarose in 100 mL TBE. Gel run for 40 mins at 110 V.