Team:DTU-Denmark/Notebook/28 June 2013

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(Created page with "=208 lab= == Main purposes today == Make PCR with x7-polymerase. ==who were in the lab== Kristian ==Procedure== Made PCRs on pZA21, GFP SF TAT, GFP SF Sec, RFP all samples...")
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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.
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.
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[[File:29.06.13 all PCR products with x7.jpg|thumb|left|The gel from the days PCR. Note the head and trail the band leaves behind it. It looks like supercoiled plasmid DNA]]
== Conclusion from today ==
== 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.
We might have used the wrong polymerase and the products we have gotten from previous PCRs with PHUSION-polymerase are without any uracil insertion.

Revision as of 01:02, 1 July 2013

Contents

208 lab

Main purposes today

Make PCR with x7-polymerase.


who were 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 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.

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