User:Eduardo Vladimir Munoz/Notebook/UNAM Genomics Mexico 2011/2011/09/16

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High Stabilitiy Plasmid Development

Abstract

  • Dephosphatation of DD B0015 E,X in pSB1AK3
  • Ligation of repC-PCR and B0015 dephosphated


Dephosphatation

The reaction was incubated at 37°C and inactivated for 5 minutes at 65°C.

Reactive Volume
DD E,X B0015 in pSB1AK3 stock 1 17μL
Antarctic Phosphatase 1μL
Reaction Buffer 10x 2μL
Total 20μL


Ligation

Reactive Volume
DD E,S repC 4μL
DD E,X B0015 in pSB1AK3 dephosph. 3μL
T4 DNA ligase 0.5μL
5x buffer 4μL
H2O mQ 8.5μL
Total 20μL



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