User:Daniela A. Garcia S./Notebook/Modeling UNAM-Genomics Mexico/2010/08/24

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WetLab: LovTap + promoters

Transformation

The ligations from 230810 were transformed into DH5α competent cells following the next protocol.

  • 20' on ice
  • 1' @ 42ºC
  • 5' on ice


  • 1h 1mL of LB
  • 1' @ 3000rmp

As a positive control was used the plasmid N.19[Am]

Ligation

A new set of ligations were done, this time with the colony_9 of LovTap. The same protocol was followed.

Ligation Protocol

  • DNA(LovTap) ..............6μL
  • Plasmid (promoter) .....2μL
  • Buffer 10X T4 Ligase ...2μL
  • Enzyme T4 Ligase .....1μL
  • HPLC ........................9μL

TOTAL 20μL

The mixture were left ON @ 16ºC



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