Difference between revisions of "Jeff Tabor/MEGAWHOP"

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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...)
 
 
Line 1: Line 1:
== PCR setup ==
+
== 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 13:46, 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