User:Meng Xiao He/Notebook/fall08/2008/11/19: Difference between revisions

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* cycles: 50
* cycles: 50
* DNA (old minipreps): 2ul each
* DNA (old minipreps): 2ul each
* for control: TOPO A


==Confirmation of pure cultures==
==Confirmation of pure cultures==

Revision as of 16:10, 19 November 2008

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One primer Phusion PCR of suicide vectors

  • extension time: 4:30
  • annealing temp: 52C
  • primer: 1ul (in 25ul of rx) of M13F primer from TOPO ki
  • cycles: 50
  • DNA (old minipreps): 2ul each
  • for control: TOPO A

Confirmation of pure cultures

  • PCR new TOPOA, pER-cco, pER-cbb
  • colony PCR of TOPO C plate

Sequencing

  • pER-Duo

Loss of plasmid

  • Streak out cultures of WM3064 on LB-agar, inoculate fresh LB liquid culture
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