User:Behzad Damadzadeh/Notebook/PcTF Subcloning in E-coli/2012/04/11

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> Digests (Fermentas FD)
BD-112 and BD-113

White dashed lines border show the vector backbone and the BD-111 constructed gene(RBS + PcTF).
Plasmid DNA 2.0 μl*
EcoR1 1.0 μl
Pst1 1.0 μl
10x FastDigest buffer + green loading dye 1.5 μl
dH2O 9.5 μl
  15.0 μl total
Incubate at 37°C for 10 minutes.
  1. Make the (1%) agarose glee and add 15μl of restricted DNA (PcTF & RBS) in one well and 10 μl of ladder.

'The size of the constructed gene supposed to be: 1123 + 15 + 6 = 1144 bp'