User:Nelson Augusto Berrocal/Notebook/WiFi Coli 2010 Wet Lab/2010/06/07

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June 07th, 2010
Because we failed the last transformation, we decided to make a PCR with the biobrick BBa_K098010 which is contained in the plasmid pSB4C5, using the preffix and suffix as primers in order to amplify only the biobrick and not the whole plasmid.

We used the next protocol:

5μL Buffer PCR

2.5μL MgCl2

2.5μL DNTPs

2.5μL Primer Forward(Preffix)

2.5μL Primer Reverse(Suffix)

4μL template DNA

1μL Taq

H2O  Total: 50μL

I made a stock for 4 samples, and finally I use three samples: a negative control lacking DNA; a positive control with 1μL of DNA and the sample to amplify with 4μL template DNA.

I made an agarose gel to .8%.

1.5 Loading Buffer

At the end of the day I analyzed the agarose gel and it seems there is any DNA, although the positive control was OK.

Ladder 1μL | C+ 5μL | C- 5μL | DNA temp 5μL | Ladder 1μL.


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