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

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04/04/11

  • ✓ Assemblies: BD111, BD112, BD113

Assemblies

  1. BD111: KAH109/(X/P)/1123 + BBa_B0030/(S/P)/15
  2. BD112: BD111/(X/P)/1138 + BBa_I712074/(S/P)46
  3. BD113: BD111/(X/P)/1138 + BBa_I719005/(S/P)23


> Digests (Fermentas FD)

Reagent Volume  
DNA (plasmid) up to 20 μL
10x buffer 3.0
enzyme 1 1.0
enzyme 2 1.0
dH2O ---
  30 μL --> 37°C/ ~30 min.
Plasmid DNA 15.0 μl*
Fermentas FastDigest enzyme 1 1.0 μl
Fermentas FastDigest enzyme 2 1.0 μl
10x FastDigest buffer + green loading dye 3.0 μl
dH2O 10.0 μl
  30.0 μl total
*For low yield DNA, use up to 25 μL; decrease dH2O accordingly.
Mix the reaction(s) thoroughly by flicking the tube.
Incubate at 37°C for 10 minutes.
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