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

Revision as of 00:10, 4 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
-*pUC19 + each flank
-*pUC19 neg control
-*4ul per transformation
-'''pir+ electrocomp cells''', LB spread w/ Gm+Xgal:
-*each pER21 prep + each flank
-*pER21 neg control
-===Rx===
-*8ul DNA mix (2:6), 1ul ligase, 1ul buffer
-*20 min ligation
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