Team:Panama/20 October 2010
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[[Image:Cuadro20octubre1.JPG|200px|thumb|left|alt text]] | [[Image:Cuadro20octubre1.JPG|200px|thumb|left|alt text]] | ||
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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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+ | [[Image:UntitledGel20Octubre1.JPG|200px|thumb|left|alt text]] | ||
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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 | Quick Check: Transformation of E. Coli with pGEM-T +Rh1ab | ||
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Electrophoresis: Quick Check | Electrophoresis: Quick Check | ||
0.8% agarose, with 1x TBE, 1.5 ul EtBr | 0.8% agarose, with 1x TBE, 1.5 ul EtBr | ||
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Samples:4, 14, 13, 22, 8, 6, 16, 12, 9, 25, 5, 1 (C-), 15, 27, 28, 10, 3, 31, 29, 11 | 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 | Homemade minipreps of Samples 1, 3, 5, 6, 10, 11, 12, 13, 16, 31 | ||
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3 ul of PCR product | 3 ul of PCR product | ||
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{{:Team:Panama/calendar2}} | {{:Team:Panama/calendar2}} |
Latest revision as of 03:54, 28 October 2010
October 20
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
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
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
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