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Salient

Salient is an excellent design with a fresh approach for the ever-changing Web. Integrated with Gantry 5, it is infinitely customizable, incredibly powerful, and remarkably simple.

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BP reaction protocol

  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
1µl
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 (pDonr201/221) or Spectinomycin (pDonr223).

Buffers

original BP buffer (5x)

  • 125 mM Tris HCl pH 7.5
  • 25 mM EDTA
  • 110 mM NaCl
  • 25 mM Spermidine-HCl
  • 5 mg/ml BSA

Note: BSA should be NON-acetylated, as Int is rapidly inactivated (at least in its pure form) by acetylated BSA

BP3 buffer (5x) - only works with attB1.1 and attB2.1

  • 100 mM Tris pH 7.5
  • 20 mM EDTA
  • 225 mM NaCl
  • 30 mM Spermidine-HCl
  • 25% Glycerol