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Team:DTU-Denmark/Notebook/28 July 2013
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| - | =lab | + | =lab 208= |
<hr/> | <hr/> | ||
==Main purpose== | ==Main purpose== | ||
<hr/> | <hr/> | ||
| + | New PCR on PAO1 for extraction of Nir - again again. | ||
| + | USER reaction between pZA21::RFP no promoter USER fragment and the araBAD USER fragment | ||
| + | PCR of pZE21 to switch out the native promoter with araBAD | ||
| + | PCR on HAO | ||
==Who was in the lab== | ==Who was in the lab== | ||
<hr/> | <hr/> | ||
| + | Kristian | ||
==Procedure== | ==Procedure== | ||
<hr/> | <hr/> | ||
| + | ===PCR=== | ||
| + | *pZE21 no promoter with primer pair 13, 57C and 3:00 | ||
| + | *HAO with primer pair 18, 59C and 3:00 | ||
| + | *Nir by colony PCR on the 4 new plates with PAO1. Primer pair 41 and 42 where used, both on [[https://2013.igem.org/Team:DTU-Denmark/Methods/PCR-touchdown| touchdown PCR]] 71→61 1C increment 10 cycles and then additional 25 cycles on 61C. | ||
| + | |||
| + | ===USER-reaction=== | ||
| + | With pZA21::RFP no promoter and the araBAD USER fragment plus two controls; one with araBAD fragment only and one with pZA21::RFP fragment only. | ||
==Results== | ==Results== | ||
Revision as of 21:12, 28 July 2013
28 July 2013
Contents |
lab 208
Main purpose
New PCR on PAO1 for extraction of Nir - again again. USER reaction between pZA21::RFP no promoter USER fragment and the araBAD USER fragment PCR of pZE21 to switch out the native promoter with araBAD PCR on HAO
Who was in the lab
Kristian
Procedure
PCR
- pZE21 no promoter with primer pair 13, 57C and 3:00
- HAO with primer pair 18, 59C and 3:00
- Nir by colony PCR on the 4 new plates with PAO1. Primer pair 41 and 42 where used, both on [touchdown PCR] 71→61 1C increment 10 cycles and then additional 25 cycles on 61C.
USER-reaction
With pZA21::RFP no promoter and the araBAD USER fragment plus two controls; one with araBAD fragment only and one with pZA21::RFP fragment only.
Results
Conclusion
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