User:Meng Xiao He/Notebook/fall08/2008/11/03: Difference between revisions

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'''DH5a''', Kan+Xgal:
'''DH5a''', Kan+Xgal:
*TOPO + each flank's original PCR
*TOPO + each flank's original PCR
'TOP10', Amp/Carb+Xgal:
'''TOP10''', Amp/Carb+Xgal:
*pUC19 + each flank
*pUC19 + each flank


'pir+ electrocomp cells', LB spread w/ Gm+Xgal:
'''pir+ electrocomp cells''', LB spread w/ Gm+Xgal:
*each pER21 prep + each flank
*each pER21 prep + each flank
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Revision as of 21:16, 1 November 2008

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Ligations/transformations

  • XmaI heat killed
  • vectors treated with ant. phosphatase
  • phosphatase killed
  • fragments column purified
  • used NEB T4 ligase b/c quick ligases have PEG

DH5a, Kan+Xgal:

  • TOPO + each flank's original PCR

TOP10, Amp/Carb+Xgal:

  • pUC19 + each flank

pir+ electrocomp cells, LB spread w/ Gm+Xgal:

  • each pER21 prep + each flank
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