User:Howard Boland/Notebook/Art from Synthetic Biology/2010/10/27
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2% Gel PCR pBR322 (790bp)
Using the new optimised conditions I ran a 2% Gel to check if I got the pBR322 product and what background noise yield it produced.
1% Gel PCR pSense66 (2302bp)
Using the new optimised conditions I ran a 1% Gel to check if I got the pSense66 product and what background noise yield it produced.
I setup a 10μl ligation between the 700bp and the 2300bp product.
Incubate for 37°C for 10 minutes.
The ligated product was heat-shock transformed and I also transformed 1μL of the sequenced pUA66katE plasmid.
This protocol is a review to correct the PCR conditions from Monday.
Mastermix PCR - pUA66katE
For each reaction add the following to each 50µl PCR tube
Cycles: 30x Lid: 100ºC Volume: 50µl (each)