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Team:SDU-Denmark/labwork/Protocols/3A
From 2012.igem.org
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<li>Digest step for Plasmid Backbone</li> | <li>Digest step for Plasmid Backbone</li> | ||
| - | + | </br> | |
<b>if sample comes from miniprep</b></br> | <b>if sample comes from miniprep</b></br> | ||
Enzyme Master Mix for Plasmid Backbone (25ul total) | Enzyme Master Mix for Plasmid Backbone (25ul total) | ||
Revision as of 21:47, 15 September 2012
| mRNA Isolation | PCR | Miniprep | Check Digest |
| 3A-Assembly | Content | Content | Content |
| Content | Content | Content | Content |
3A-Assembly -Ligation of 2 parts into a vector
- PCR step; see PCR protocol
- Digestion BSA is only needed for conventional restriction enzymes, NOT fast digest
- Digest step for Plasmid Backbone if sample comes from miniprep Enzyme Master Mix for Plasmid Backbone (25ul total)
- 5 ul Fastdigest greenbuffer / NEB Buffer 2
- 0.5 ul BSA
- 0.5 ul EcoRI-H
- 0.5 ul PstI
- 19 ul dH20 (0,5 ul less when using BSA)
