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Protocol for Blunting DNA with 5'- or 3'- protruding Termini
(with T4 DNA Polymerase)

1. Prepare the following reaction mixture:
   

   5X reaction buffer for T4 DNA Polymerase	4 µl
   digested DNA or PCR product	1 µg
   dNTP Mix, 2 mM each	1 µl (0.1 mM final concentration)
   T4 DNA Polymerase	0.2 µl (1 u)
   Water, nuclease-free	to 20 µl

2. Incubate the mixture at 11°C for 20 minutes or at room temperature for 5 minutes.
3. Stop the reaction by heating at 75°C for 10 minutes.

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. Ausubel, F.M., et al., Current Protocols in Molecular Biology, vol. 1, John Wiley & Sons, Inc., Brooklyn, New York, 3.5.11-3.5.12, 1994-2004.

Ordering Information

T4 DNA Polymerase dNTP Mixes dNTP Set T4 DNA Ligase 0.5 M EDTA, pH 8.0 Water, nuclease-free

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Updated gegužės 09, 2007 17:26