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Contents 1 208 lab 1.1 Main purposes today 1.2 Who was in the lab 1.3 Procedure 1.4 Results 1.5 Conclusion from today

208 lab

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Main purposes today

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Make PCR with x7-polymerase.

Who was in the lab

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Kristian

Procedure

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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

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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

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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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