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Team:DTU-Denmark/Notebook/28 June 2013
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=208 lab= | =208 lab= | ||
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== Main purposes today == | == Main purposes today == | ||
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Make PCR with x7-polymerase. | Make PCR with x7-polymerase. | ||
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==who were in the lab== | ==who were in the lab== | ||
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Kristian | Kristian | ||
==Procedure== | ==Procedure== | ||
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Made PCRs on pZA21, GFP SF TAT, GFP SF Sec, RFP all samples where made in duplicates. | Made PCRs on pZA21, GFP SF TAT, GFP SF Sec, RFP all samples where made in duplicates. | ||
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Second program was 68°C annealing and 2:00 extension time with tube: 9+10, 11+12, 13+14, 15+16. | 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. | Last program was 70°C annealing and 1:00 extension time with tube: 17+18. | ||
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==Results== | ==Results== | ||
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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]] <html><p> <br/> </p></html>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. | |
| - | [[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]] | + | <html><p> <br/><br/><br/><br/><br/><br/><br/> </p></html> |
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== Conclusion from today == | == 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. | 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 08:06, 12 July 2013
28 June 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
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.
