IGEM:Slovenia HS 2015/2009/Notebook/Test/2015/07/21

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CtfA, CtfB, BdhB restriction

Mixture:

  • 20 μL DNA
  • 2,5 μL buffer Tango
  • 1 μL Xba1
  • 2 μL Psrt1

Restriction mixture was incubated at 37°C for 3h.

PCR

Markings: 1-10 CtfA, 11-20 CtfB, 21-30 BdhB

Agarose gel electrophoresis

Expected lenghts of genes:

  • CtfA: 735 bp
  • CtfB: 744 bp
  • BdhA: 1250 bp
  • AdhE1: 1666 bp
  • AdhE2: 1020 BP

Electrophoresis was successful for the restriction products. Fragments with appropriate lenghts were cut from the gel.

DNA isolation from agarose gel

  • protocol- manjka še!

Ligation

po protokolu- manjka še!

  • overnight ligation

These fragments were then inserted into the expression vector pSB1C3.



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