User:Nelson Augusto Berrocal/Notebook/WiFi Coli 2010 Wet Lab/2010/06/22/2010/06/23

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June 23rd, 2010

I made a PCR with Rtth to ampliffy LuxAB from Vibrio Fischeri MJ11 and obtain more product in order to purify it; I made four reactions following the next protocol (five reaction counting the control):
1st solution
2μL (two reactions) and 1μL (two reactions) template DNA (Genomic DNA)
6μL Bufer 3.3x
3μL Mg(Ac)2
4μL dNTPs
3μL Forward Primer
3μL Reverse Primer
10μL H2O
30μL TOTAL

After a hotstart I added the 2nd solution
10.5μL H2O
9μL Buffer 3.3x
0.5μL Rtth polymerase
TOTAL 20μL

The four samples and control were amplified.

We also received E.coli K12 EnvZ mutant from Princeton and I cultured it in a LB petri box.

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