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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