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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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LR reaction protocol

  1. Assemble the reaction:
5 µl
10 µl
Entry clone (50-150ng)
x µl
x µl
Destination vector
75 ng
150 ng
5X LR Buffer
1 µl
2 µl
TE pH 8.0
to 4 µl
to 8 µl
LR Clonase
0.5µl
1 µl
  1. Incubate at 25°C for 1 hour to overnight.
  2. Transform into competent cells and plate on plates containing a suitable antibiotic.

Buffers

Original LR buffer (5x)

  • 250 mM of Tris HCl pH 7.5
  • 2.25 mM of EDTA
  • 250 mM of NaCl
  • 12.5 mM of spermidine HCl
  • 1 mg/mL of BSA

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

LR4 buffer (5x) - for use with attL1.1 and attL2.1

  • 200 mM Tris-Cl, pH 7.5
  • 10 mM EDTA
  • 320 mM NaCl
  • 35 mM spermidine-HCl
  • 25% Glycerol