Jeff Tabor/MEGAWHOP: Difference between revisions
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(New page: == PCR setup == *120ng template plasmid *500ng Megaprimer *Pfu Ultra II == Cycling conditions == 95C, 2' <br> 95C, 20" <br> 55C, 20" <br> 68C, 6-12' <br> 25x <br> == Transformation == *D...) |
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== | == Reaction setup == | ||
*120ng template plasmid | *120ng template plasmid | ||
*500ng Megaprimer | *500ng Megaprimer | ||
| Line 13: | Line 13: | ||
== Transformation == | == Transformation == | ||
*DpnI, 1-2h <br> | *DpnI, 1-2h <br> | ||
*Transform 0.5-2uL into 50uL chemically competent ''E.coli'' | *Qiaclean the reaction <br> | ||
*Transform 0.5-2uL clean DNA into 50uL chemically competent ''E.coli'' | |||
*Plate all | |||
Latest revision as of 14:46, 13 December 2010
Reaction setup
- 120ng template plasmid
- 500ng Megaprimer
- Pfu Ultra II
Cycling conditions
95C, 2'
95C, 20"
55C, 20"
68C, 6-12'
25x
Transformation
- DpnI, 1-2h
- Qiaclean the reaction
- Transform 0.5-2uL clean DNA into 50uL chemically competent E.coli
- Plate all
