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Linker Ligation
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Linker Ligation (with T4 DNA Ligase)

  1. In a microcentrifuge tube prepare a solution of blunt ended, dephosphorylated DNA (100-500ng) in TE buffer (5-7µl).
  2. Add 1-2µg of phosphorylated linkers in 5µl of TE buffer.
  3. Add:
  4. Incubate the mixture for 1 hour at 22°C .
  5. Inactivate T4 DNA Ligase by heating the reaction mixture at 65°C for 10 minutes. The resulting ligation products can be digested directly with restriction endonucleases.

 Note 

Reference

Wu, R. et al., Meth. Enzymol., 152, 343, 1987.

Ordering Information

T4 DNA Ligase #EL0015
#EL0016
#EL0014
#EL0011
#EL0012
#EL0017
#EL0013		 LC, 200u (1u/µl)
LC, 5x200u (1u/µl)
200u (5u/µl)
1000u (5u/µl)
5x100u (5u/µl)
HC, 1000u (30u/µl)
HC, 5000u (30u/µl)
T4 DNA Ligase (with PEG) #EL0335
#EL0336
#EL0334
#EL0331
#EL0337		 LC, 200u (1u/µl)
LC, 5x200u (1u/µl)
200u (5u/µl)
1000u (5u/µl)
HC, 1000u (30u/µl)
Rapid DNA Ligation Kit #K1421
#K1422		 for 10 reactions
for 50 reactions
Rapid DNA Ligation & Transformation Kit #K1431
#K1432		 for 10 reactions
for 30 reactions
DNA Blunting & Ligation Kit #K1511
#K1512		 for 10 reactions
for 30 reactions
DNA Cloning Kit #K1311
#K1312		 for 10 reactions
for 30 reactions
InsT/Aclone™ PCR Product Cloning Kit #K1213
#K1214		 for 10 reactions
for 30 reactions
TransformAid™ Bacterial Transformation Kit #K2710
#K2711		 for 20 transformations
for 40 transformations
Dovetail™ PCR Product Cloning Kit #K1911
#K1912		 for 10 reactions
for 30 reactions
MIDGE® Vector Production Kit #K2901 for 6 MIDGE® vectors
N.Bpu10I #ER1681
#ER1682		 100u
500u
T7 DNA Polymerase #EP0081 300u (10u/µl)
DEPC-treated Water, molecular biology grade #R0601 30ml

 

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Updated August 23, 2002 13:51