IGEM:Paris Bettencourt 2012/Notebooks/Semantic group/day by day//2012/10/16: Difference between revisions
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I plated everything on LB+Amp, and let it grow O/N at 37°C. | I plated everything on LB+Amp, and let it grow O/N at 37°C. | ||
==Soil sample analysis== | |||
I took 3 sample of soil from the sidewalk, at 3 different places, that weight about 20-30g. I add 25mL of distilled water and vortex hard for couples of minutes. I let settled over night. | |||
This morning I took in duplicate 50µL, 25µL, 15µL, 5µL of supernatant for sample. I add water up to 50µL. I put this at 99°C for 10min, then I spin it down for 5min, and I took 5µL of the supernatant. | |||
So, I ran the PCR with : | |||
• 5µL of supernatant | |||
• 15µL of water | |||
• 2,5µL of primers (VF2 & VR) | |||
• 25µL of PCR MM (contain taq, dNTP, ..) | |||
PCR pgm : | |||
95°C - 1' | |||
95°C - 30" | |||
50°C - 30" | |||
72°C - 1' | |||
72°C - 10' | |||
4°C -\infty | |||
==Soil sample test== | ==Soil sample test== |
Revision as of 12:14, 16 October 2012
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PCR QCM - 2nd part
I plated everything on LB+Amp, and let it grow O/N at 37°C. Soil sample analysisI took 3 sample of soil from the sidewalk, at 3 different places, that weight about 20-30g. I add 25mL of distilled water and vortex hard for couples of minutes. I let settled over night. This morning I took in duplicate 50µL, 25µL, 15µL, 5µL of supernatant for sample. I add water up to 50µL. I put this at 99°C for 10min, then I spin it down for 5min, and I took 5µL of the supernatant. So, I ran the PCR with : • 5µL of supernatant • 15µL of water • 2,5µL of primers (VF2 & VR) • 25µL of PCR MM (contain taq, dNTP, ..) PCR pgm : 95°C - 1' 95°C - 30" 50°C - 30" 72°C - 1' 72°C - 10' 4°C -\infty Soil sample test |