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Team:DTU-Denmark/Notebook/2 August 2013
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==Procedure== | ==Procedure== | ||
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| + | ===Gel purification=== | ||
* Gel purification was performed according to protocol included in QIAEX, Gel Extraction Kit. | * Gel purification was performed according to protocol included in QIAEX, Gel Extraction Kit. | ||
| + | ===USER reaction=== | ||
* USER reaction was performed according to standard protocol SOP1. We use DNA linear fragment containing pZA21 bacbone,cycAX with His-Tag sequence. | * USER reaction was performed according to standard protocol SOP1. We use DNA linear fragment containing pZA21 bacbone,cycAX with His-Tag sequence. | ||
| + | ===PCR === | ||
* PCR according to standard PCR protocol. | * PCR according to standard PCR protocol. | ||
Samples names: | Samples names: | ||
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* 9, 10 - Tat His-Tag, primers 20a, 20b; template: TAT 3 1a | * 9, 10 - Tat His-Tag, primers 20a, 20b; template: TAT 3 1a | ||
* 11, 12, 13 - negative controls as follows for: Tat His-Tag, Nir, Sec His-Tag | * 11, 12, 13 - negative controls as follows for: Tat His-Tag, Nir, Sec His-Tag | ||
| - | + | ===Purification of plasmids after ethanol purification=== | |
| + | According to protocol attached to the DNA purification kit, QIAGEN | ||
==Results== | ==Results== | ||
<hr/> | <hr/> | ||
Revision as of 15:22, 2 August 2013
2 August 2013
Contents |
lab 208
Main purpose
- Gel purification of Cyc and Nir
- USER reaction
- PCR of Nir, His-Tag Sec, His-Tag TAT
- Miniprep re-purification with columns
Who was in the lab
Gosia, Kristian, Natalia, Julia
Procedure
Gel purification
- Gel purification was performed according to protocol included in QIAEX, Gel Extraction Kit.
USER reaction
- USER reaction was performed according to standard protocol SOP1. We use DNA linear fragment containing pZA21 bacbone,cycAX with His-Tag sequence.
PCR
- PCR according to standard PCR protocol.
Samples names:
- 1,2 - Nir 48 (primers 48a, 48b) on template of Nir (too long fragment amplified at the beginning of work with Nir PCR)
- 3,4 - Sec His-Tag, primers 19a, 19b; template: Sec 2
- 5,6 - Tat His-Tag, primers 20a, 20b; template: TAT 2 1
- 7,8 - Tat His-Tag, primers 20a, 20b; template: TAT 3 2
- 9, 10 - Tat His-Tag, primers 20a, 20b; template: TAT 3 1a
- 11, 12, 13 - negative controls as follows for: Tat His-Tag, Nir, Sec His-Tag
Purification of plasmids after ethanol purification
According to protocol attached to the DNA purification kit, QIAGEN
Results
Conclusion
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