Team:HKUST/Protocols/gel cut
From 2009.igem.org
(Difference between revisions)
Line 44: | Line 44: | ||
<ul> | <ul> | ||
<li><a href="https://2009.igem.org/Team:Gallery">Gallery</a></li> | <li><a href="https://2009.igem.org/Team:Gallery">Gallery</a></li> | ||
- | <li><a href="https://2009.igem.org/Team: | + | <li><a href="https://2009.igem.org/Team:Biosafety">Biosafety</a></li> |
<li><a href="https://2009.igem.org/Team:Acknowledgement">Acknowledgement</a></li> | <li><a href="https://2009.igem.org/Team:Acknowledgement">Acknowledgement</a></li> | ||
</ul> | </ul> |
Revision as of 16:48, 19 October 2009
a
Cutting bands on the gel
Purpose: To extract specific DNA fragment.
Procedure:
a.Visualize the bands under UV light. Use long-wavelength UV to minimize damage to the DNA.b.Cut the band with a clean razor blade.
c.Turn the gel slice on its side to trim off extra agarose. Place the gel in a microcentrifuge tube.
Tips:
EB staining is needed for higher resolution under UV light.Safety tips:
1. ermission is needed from technician before doing this! Follow the safety instruction in the room.2.When using UV light, protect your skin by wearing safety goggles or a face shield, gloves, and a lab coat.
- Agarose gel preparation and gel electrophoresis
- PCR (Taq, Vent)
- PCR product clean-up
- Enzyme digestion (vector, insert)
- Plasmid DNA extraction
- Gel cutting
- Gel extraction
- Ethanol precipitation
- Ligation
- Transformation to E.coli
- Transformation to yeast
- Cell lysis
- Yeast genomic DNA extraction
- Western blotting
- GFP testing