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Team:DTU-Denmark/Notebook/25 July 2013
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===ON cultures and re-plating of Nir USER transformants from Colony PCR=== | ===ON cultures and re-plating of Nir USER transformants from Colony PCR=== | ||
| - | Positive Nir USER transformants obtained by colony PCR | + | Positive Nir USER transformants obtained by colony PCR: |
| - | Samples 25 and 26 were inoculated in 5 mL LB medium + 30 ug/ml kanamicyn for ON cultures preparation. | + | * Samples 25 and 26 were inoculated in 5 mL LB medium + 30 ug/ml kanamicyn for ON cultures preparation. |
| - | Samples 25, 26 and 27 were re-plated in LB agar + 30 ug/ml kanamicyn | + | * Samples 25, 26 and 27 were re-plated in LB agar + 30 ug/ml kanamicyn |
==Results== | ==Results== | ||
Revision as of 12:57, 25 July 2013
Contents |
208
Main purpose
- PCR of AMO and HAO using USER primers and as a template using AMO and HAO extracted from chromosomal DNA with non-uracil primers
- verification of PCRs
- ON cultures and re-plating of Nir USER transformants from Colony PCR ( 17-07-2013)
Who was in the lab
Kristian, Gosia, Julia
Procedure
PCR in order to amplify AMO and HAO with USER primers
gel electrophoresis
ON cultures and re-plating of Nir USER transformants from Colony PCR
Positive Nir USER transformants obtained by colony PCR:
- Samples 25 and 26 were inoculated in 5 mL LB medium + 30 ug/ml kanamicyn for ON cultures preparation.
- Samples 25, 26 and 27 were re-plated in LB agar + 30 ug/ml kanamicyn
Results
Gel 1
- 1 kb ladder
- AMO 5uL template, 1
- AMO 5uL template, 2
- AMO 10uL template, 1
- AMO 10uL template, 2
- HAO 5uL template, 1
- HAO 5uL template, 2
- HAO 10uL template, 1
- HAO 10uL template, 2
- restriction analysis cyc EcoRI
- restriction analysis cyc HindIII
- 1 kb ladder
Gel 2
Conclusion
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