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

From OpenWetWare
Jump to: navigation, search
Owwnotebook icon.png Project name Report.pngMain project page

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

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

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.