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Team:SDU-Denmark/labwork/Protocols/3A
From 2012.igem.org
(Difference between revisions)
| Line 282: | Line 282: | ||
<li>Digestion</li> | <li>Digestion</li> | ||
<i>BSA is only needed for conventional restriction enzymes, NOT fast digest</i> | <i>BSA is only needed for conventional restriction enzymes, NOT fast digest</i> | ||
| + | <ol type="i"> | ||
| + | <li>Digest step for Plasmid Backbone</li> | ||
<ul type="square"> | <ul type="square"> | ||
| - | |||
| - | |||
<b>if sample comes from miniprep</b> | <b>if sample comes from miniprep</b> | ||
| + | <li>Enzyme Master Mix for Plasmid Backbone (25ul tota</li> | ||
| + | <ul> | ||
| + | <li>5 ul Fastdigest greenbuffer / NEB Buffer 2</li> | ||
| + | <li>0.5 ul BSA</li> | ||
| + | <li>0.5 ul EcoRI-H</li> | ||
| + | <li>0.5 ul PstI</li> | ||
| + | <li>19 ul dH20 (0,5 ul less when using BSA)</li> | ||
| + | |||
| + | </ul> | ||
</ul> | </ul> | ||
</ol> | </ol> | ||
| - | + | ||
<li></li> | <li></li> | ||
<li></li> | <li></li> | ||
Revision as of 21:45, 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
- Enzyme Master Mix for Plasmid Backbone (25ul tota
- 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)
-
if sample comes from miniprep
