"
Team:UNIPV-Pavia/Protocols/PlasmidExtraction
From 2008.igem.org
(Difference between revisions)
| (11 intermediate revisions not shown) | |||
| Line 14: | Line 14: | ||
<br> | <br> | ||
| + | |||
| + | <h1>The protocols we used</h1> | ||
| + | |||
| + | *[[Team:UNIPV-Pavia/Protocols/Lb|LB medium preparation]] | ||
| + | *[[Team:UNIPV-Pavia/Protocols/Resuspension|Plasmid resuspension from IGEM paper spots]] | ||
| + | *[[Team:UNIPV-Pavia/Protocols/Transformation|Transformation]] | ||
| + | *[[Team:UNIPV-Pavia/Protocols/PlasmidExtraction|Plasmid extraction]] | ||
| + | *[[Team:UNIPV-Pavia/Protocols/Digestion|BioBrick digestion with restriction enzymes]] | ||
| + | *[[Team:UNIPV-Pavia/Protocols/GelExtraction|DNA gel extraction]] | ||
| + | *[[Team:UNIPV-Pavia/Protocols/Precipitation|DNA precipitation with sodium acetate]] | ||
| + | *[[Team:UNIPV-Pavia/Protocols/AntarcticPhosphatase|Antarctic Phosphatase]] | ||
| + | *[[Team:UNIPV-Pavia/Protocols/Ligation|Ligation]] | ||
| + | *[[Team:UNIPV-Pavia/Protocols/Pcr|PCR]] | ||
| + | |||
<h1>Plasmid extraction</h1> | <h1>Plasmid extraction</h1> | ||
| - | + | ''(Estimated time: 3 hours if you use QIAGEN Plasmid Mini Kit; 1 hour if you use QIAGEN QIAprep Spin Miniprep Kit)'' | |
| - | ''( | + | |
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
| - | + | ||
<br> | <br> | ||
<br> | <br> | ||
'''Materials needed:''' | '''Materials needed:''' | ||
| - | *''' | + | *'''5 ml or 9 ml (for higher yields) overnight mini-culture |
| - | *'''QIAGEN QIAprep Spin | + | *'''QIAGEN Plasmid Mini Kit or QIAGEN QIAprep Spin Miniprep Kit''' |
<br> | <br> | ||
| - | * | + | *Follow QIAGEN standard protocol: [http://www1.qiagen.com/literature/render.aspx?id=369&tp=4 QIAGEN Plasmid Mini Kit protocol] or [http://www1.qiagen.com/literature/render.aspx?id=370&tp=4 QIAGEN QIAprep Spin Miniprep protocol] |
| + | *We used QIAGEN Plasmid Mini Kit in the first 2 weeks of our activity to perform miniprep, when we didn't have QIAGEN QIAprep Spin Miniprep Kit at our disposal. Then, we always used QIAGEN QIAprep Spin Miniprep Kit for its higher yield. | ||
<br> | <br> | ||
Latest revision as of 12:52, 1 July 2008
 Home
|  The Team
|  The Project
|  Biological Safety
|  Parts Submitted to the Registry
|
|---|---|---|---|---|
 Dry Lab
|  Wet Lab
|  Modeling
|  Protocols
|  Activity Notebook
|
The protocols we used
- LB medium preparation
- Plasmid resuspension from IGEM paper spots
- Transformation
- Plasmid extraction
- BioBrick digestion with restriction enzymes
- DNA gel extraction
- DNA precipitation with sodium acetate
- Antarctic Phosphatase
- Ligation
- PCR
Plasmid extraction
(Estimated time: 3 hours if you use QIAGEN Plasmid Mini Kit; 1 hour if you use QIAGEN QIAprep Spin Miniprep Kit)
Materials needed:
- 5 ml or 9 ml (for higher yields) overnight mini-culture
- QIAGEN Plasmid Mini Kit or QIAGEN QIAprep Spin Miniprep Kit
- Follow QIAGEN standard protocol: [http://www1.qiagen.com/literature/render.aspx?id=369&tp=4 QIAGEN Plasmid Mini Kit protocol] or [http://www1.qiagen.com/literature/render.aspx?id=370&tp=4 QIAGEN QIAprep Spin Miniprep protocol]
- We used QIAGEN Plasmid Mini Kit in the first 2 weeks of our activity to perform miniprep, when we didn't have QIAGEN QIAprep Spin Miniprep Kit at our disposal. Then, we always used QIAGEN QIAprep Spin Miniprep Kit for its higher yield.
