Team:DTU-Denmark/Notebook/28 June 2013

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28 June 2013

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Contents

208 lab


Main purposes today


Make PCR with x7-polymerase.

Who was in the lab


Kristian

Procedure


Made PCRs on pZA21, GFP SF TAT, GFP SF Sec, RFP all samples where made in duplicates.

In tube 1+2, 9+10 where pZA21. In 3+4, 11+12 GFP SF TAT In 5+6, 13+14, 17+18 GFP SF Sec In 7+8, 15+16 RFP

First program was 59°C annealing and 2:00 extension time with tube: 1+2, 3+4, 5+6, 7+8. Second program was 68°C annealing and 2:00 extension time with tube: 9+10, 11+12, 13+14, 15+16. Last program was 70°C annealing and 1:00 extension time with tube: 17+18.

Results


The gel from the days PCR. Note the head and trail the band leaves behind it. It looks like supercoiled plasmid DNA


The days PCR where not very successful. The gel pic. looks like that we can only see supercoiled plasmid DNA which must originate from the template DNA.








Conclusion from today


We might have used the wrong polymerase and the products we have gotten from previous PCRs with PHUSION-polymerase are without any uracil insertion.

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