BP reaction

  1. Assemble the reaction:
   
5 µl
10 µl
  attB1-PCR or linearized expresssion clone
x µl
x µl
  pDONR vector
75 ng
150 ng
  5X BP Buffer
1 µl
2 µl
  TE pH 8.0
to 4 µl
to 8 µl
  BP Clonase
1 µl
2 µl
  1. Incubate at 25°C for 1 hour to overnight.
  2. Add 1 or 2 µl of Proteinase K solution to each reaction. Incubate for 10 minutes at 37°C.
  3. Transform into competent cells and plate on plates containing Kanamycin.