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Self-circularization of Linear DNA
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Self-circularization of Linear DNA (with T4 DNA Ligase)

  1. In a microcentrifuge tube prepare a solution of linear DNA (25-50ng) in deionized water or TE buffer (10-35µl).
  2. Add:
  3. Incubate the mixture for 1 hour at 22°C .
  4. Inactivate T4 DNA Ligase by heating reaction mixture at 65°C for 10 minutes.
  5. Resulting reaction mixture can be used directly for transformation.

References

  1. Sambrook, J., Russell, D.W., Molecular Cloning: A Laboratory Manual, the third edition, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, 2001.
  2. Current Protocols in Molecular Biology, vol. 1 (Ausubel, F.M., et al., ed.), John Wiley & Sons, Inc., Brooklyn, New York, 1994-2001.

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