Team:Panama/20 October 2010

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iGEM Panama

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October 20

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There was a PCR done to sample #2. PCR: to amplify RH1ab, samples of plasmid DNA. We inoculated 30 colonies en 3ml liquid LB medium + Amp at 37C, overnight with vigorous shaking. 30 white colonies and one blue colony as a negative control

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Digestion of pGEM-T + Rh1AB to compare with the non-digested and verify the difference in size Note: EcoR1 releases the insert.

Quick Check: Transformation of E. Coli with pGEM-T +Rh1ab

-100 ul culture -50 ul phenol:chloroform -10 ul loading buffer

Vortex

Electrophoresis: Quick Check 0.8% agarose, with 1x TBE, 1.5 ul EtBr Samples:4, 14, 13, 22, 8, 6, 16, 12, 9, 25, 5, 1 (C-), 15, 27, 28, 10, 3, 31, 29, 11


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Homemade minipreps of Samples 1, 3, 5, 6, 10, 11, 12, 13, 16, 31

Cloning: pGEM-T + Rh1ab -Ligation: Overnight at 16C Ratio of insert:vector as 1:1

3 ul of PCR product


May
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August
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30 31
September
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October
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