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Team:SDU-Denmark/labwork/Protocols/3A
From 2012.igem.org
(Difference between revisions)
| Line 297: | Line 297: | ||
</br> | </br> | ||
<b>If sample comes from PCR</b> | <b>If sample comes from PCR</b> | ||
| + | Enzyme Master Mix for Plasmid Backbone (25ul total) | ||
| + | <ul type="square"> | ||
| - | + | <li>5 ul Fastdigest greenbuffer / NEB Buffer </li> | |
| + | <li>0.5 ul BSA</li> | ||
| + | <li>0.5 ul EcoRI-HF</li> | ||
| + | <li>0.5 ul PstI</li> | ||
| + | <li>0.5 ul DpnI (Used to digest any template DNA from productio</li> | ||
| + | <li>19 ul dH20 (0,5 ul less when using BSA</li> | ||
| + | </ul> | ||
</ol> | </ol> | ||
Revision as of 21:51, 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)
- 5 ul Fastdigest greenbuffer / NEB Buffer
- 0.5 ul BSA
- 0.5 ul EcoRI-HF
- 0.5 ul PstI
- 0.5 ul DpnI (Used to digest any template DNA from productio
- 19 ul dH20 (0,5 ul less when using BSA
