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Team:DTU-Denmark/Notebook/13 July 2013
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| - | {{:Team:DTU-Denmark/Templates/StartPage| | + | {{:Team:DTU-Denmark/Templates/StartPage|13 July 2013}} |
| - | + | Navigate to the [[Team:DTU-Denmark/Notebook/12_July_2013|Previous]] or the [[Team:DTU-Denmark/Notebook/14_July_2013|Next]] Entry | |
| - | + | ||
=208 lab= | =208 lab= | ||
<hr/> | <hr/> | ||
== Main purpose today == | == Main purpose today == | ||
<hr/> | <hr/> | ||
| + | Make gel purifications to enable USER-reactions and to make PCR of the fragments missing. | ||
==Who was in the lab== | ==Who was in the lab== | ||
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==Procedure== | ==Procedure== | ||
<hr/> | <hr/> | ||
| + | ===Purification gel 1,2 and 3=== | ||
| - | * | + | *1kb ladder |
| + | *Nir | ||
| + | *Nir | ||
| + | *Nir | ||
| + | *Nir | ||
| + | *Nir | ||
| + | *Nir | ||
| + | *Nir | ||
| + | *Nir | ||
| + | *HAO | ||
[[File:2013 07 13 gel Nir+HAO USER puri.jpg| 600px]] | [[File:2013 07 13 gel Nir+HAO USER puri.jpg| 600px]] | ||
| - | * | + | *1kb ladder |
| + | *cycAX | ||
| + | *cycAX | ||
| + | *cycAX | ||
| + | *cycAX | ||
| + | *AMO | ||
| + | *AMO | ||
| + | *AMO | ||
| + | *AMO | ||
| + | *HAO | ||
| + | *HAO | ||
| + | *HAO | ||
[[File:J10893.jpg| 600px]] | [[File:J10893.jpg| 600px]] | ||
*100 bp ladder, NEB | *100 bp ladder, NEB | ||
| + | *RFP | ||
| + | *RFP | ||
| + | *RFP | ||
[[File:This is not a fucking duplicate.jpg| 600px]] | [[File:This is not a fucking duplicate.jpg| 600px]] | ||
| + | ===PCR of AMO=== | ||
| + | Did PCR on AMO on two programs: | ||
| + | 1-3 on 54°C | ||
| + | 4-6 on 52°C | ||
| + | Both with the annealing time 3 min. | ||
| + | |||
| + | ===USER-reactions=== | ||
| + | There was made USER-reactions with all fragments after purification. AMO was purified even though it was only one very faint band; the others were bright band. The following plates where made: | ||
| + | *RFP USER - RFP inside pZA21 | ||
| + | *AMO USER - AMO inside pZA21 | ||
| + | *HAO USER - HAO inside pZA21 | ||
| + | *cycAX USER - cycAX inside pZA21 | ||
| + | *Nir USER - Nir inside pZA21 | ||
| + | *Neg. USER - pZA21 linearized only | ||
| + | |||
| + | ==Conclusion== | ||
| + | <hr/> | ||
| + | PCR of AMO with USER-primers was not a success we will see if the new PCR have done better. We will evaluate the transformants on Monday with colony-PCR. | ||
| + | |||
| + | Navigate to the [[Team:DTU-Denmark/Notebook/12_July_2013|Previous]] or the [[Team:DTU-Denmark/Notebook/14_July_2013|Next]] Entry | ||
{{:Team:DTU-Denmark/Templates/EndPage}} | {{:Team:DTU-Denmark/Templates/EndPage}} | ||
Latest revision as of 20:40, 16 September 2013
13 July 2013
Contents |
208 lab
Main purpose today
Make gel purifications to enable USER-reactions and to make PCR of the fragments missing.
Who was in the lab
Kristian
Procedure
Purification gel 1,2 and 3
- 1kb ladder
- Nir
- Nir
- Nir
- Nir
- Nir
- Nir
- Nir
- Nir
- HAO
- 1kb ladder
- cycAX
- cycAX
- cycAX
- cycAX
- AMO
- AMO
- AMO
- AMO
- HAO
- HAO
- HAO
- 100 bp ladder, NEB
- RFP
- RFP
- RFP
PCR of AMO
Did PCR on AMO on two programs: 1-3 on 54°C 4-6 on 52°C Both with the annealing time 3 min.
USER-reactions
There was made USER-reactions with all fragments after purification. AMO was purified even though it was only one very faint band; the others were bright band. The following plates where made:
- RFP USER - RFP inside pZA21
- AMO USER - AMO inside pZA21
- HAO USER - HAO inside pZA21
- cycAX USER - cycAX inside pZA21
- Nir USER - Nir inside pZA21
- Neg. USER - pZA21 linearized only
Conclusion
PCR of AMO with USER-primers was not a success we will see if the new PCR have done better. We will evaluate the transformants on Monday with colony-PCR.
Navigate to the Previous or the Next Entry
