From 2012.igem.org
Team:Technion/17_June_2012
Ilya
I had a full week planned in order to finish as much progress as possible but of course something had to go wrong. On Thursday I ran the PCR to add the restriction sites to our fusion PCR product. Following the instructions for the polymerase, I checked the annealing temperature using the NEB Ta calculator. It gave me a Ta of 69 degrees. Apparently, that was way too high. Asaf and I spent most of today running gels to figure out what happened to our PCR product.
In the first gel, Asaf used a 100 bp marker instead of a 1kb marker so we couldn't figure out what was the size of the band we saw. Then I used the same gel to run a new sample with a new 1kb marker but then the gel turned out blank. As if there was no ethidium bromid left. So I prepared a new gel and ran it. Then it was finally obvious that we didn't get a PCR product but some weird band at way less than 500 bp. After talking to Roee and Sharon we learned that the Ta should be a lot lower, despite what the NEB calculator gave us. So I set up the PCR reaction again and tomorrow we will see if we finally got a product.
Our weekly plan will be delayed for a day or two, bummer.
Inbal
Asaf
Hila
Lior
Noa
Evgeni
Shahar
Rachel
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