This is a cached page for the URL (http://www.fermentas.com/techinfo/ModifyingEnzymes/protocols/p_dnainslig.htm). To see the most recent version of this page, please click here.
Protocol Online is not affiliated with the authors of this page nor responsible for its content.
About Cache
DNA Insert Ligation into Vector DNA
 Home |Contacts |Order |Catalog |Support
 Search |Alphabetical Index |Numerical Index
Fermentas ISO9002
 Restriction Endonucleases |Modifying Enzymes |PCR |Kits |Markers  Nucleic Acids
|Nucleotides & Oligonucleotides | Media |Transfection Reagents |Reagents

S

U

P

P

O

R

T

 

DNA Insert Ligation into Vector DNA (with T4 DNA Ligase)

  1. In a microcentrifuge tube prepare 5-10µl mix in water or TE buffer of digested (if preferred dephosphorylated and purified) vector DNA (50-400ng) and foreign DNA to be inserted.
  2. Add the following components to the same tube:
  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. Use the mixture for transformation.

 Note 

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

 

 Home |Search |Contacts |Order |Catalog |Support

catalog@fermentas.com

Updated August 23, 2002 13:51