User:Behzad Damadzadeh/Notebook/PcTF Subcloning in E-coli/2013/09/07

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09/06/2013

Confirm The Mutagenesis

Plasmid Type SD-BD0021 SD-BD002 SD-BD002 BD002 (-) Ctrl BD002 (+)Ctrl BD002 (+) Ctrl
Plasmid DNA 3.0 μl 3.0 μl 3.0 μl 3.0 μl 3.0 μl 3.0 μl
BsmBI 1.0 μl 0.0 μl 0.0 μl 1.0 μl 0.0 μl 0.0 μl
Spe1 0.0 μl 1.0 μl 0.0 μl 0.0 μl 1.0 μl 0.0 μl
10x FastDigest buffer + green loading dye 1.5 μl 1.5 μl 1.5 μl 1.5 μl 1.5 μl 1.5 μl
dH2O 9.5 μl 9.5 μl 10.5 μl 9.5 μl 9.5 μl 10.5 μl
  15.0 μl total 15.0 μl total 15.0 μl total 15.0 μl total 15.0 μl total 15.0 μl total
Incubate at 37°C for 10 minutes.

Ste-Directed-Test.jpg

  1. Make the (1%) agarose glee and add 15μl of restricted vector in one well and 10 μl of ladder.
  • The gel picture shows that the BSMBI cut site did not remove.