- Preparation of blunt-end DNA fragments (Practical Approach Online, Oxford University Press)
Two methods are introduced: Filling in 5'-overhangs with Klenow DNA polymerase and removal of 3'-overhangs by the exonuclease reaction of T4
Added: Sat Aug 17 2002, Hits: 2039, Reviews: 0
Applications of Modifying Enzymes
Presents quick reference table for application of modifying enzymes.
Added: Tue May 14 2002, Hits: 81, Reviews: 0
Electroelution of DNA Fragments From Agarose into Dialysis Tubing
(Donis Keller Lab)
To retrieve and purify any specific DNA fragment from an agarose gel slice; the expected yield is from 50-75% of the amount in the gel slice.
Added: Tue May 14 2002, Hits: 230, Reviews: 0
(Hedrick Lab, UCSD Cancer Center)
Reaction for generating blunt ends on dsDNA fragments with overhangs (especially good for 5' overhangs).
Added: Tue Dec 10 2002, Hits: 403, Reviews: 0
A simple, reversible way to a stop restriction reaction is by adding EDTA, which chelates Mg2+, thereby preventing catalysis. If further manipulations of the digested DNA are to be performed, the restriction endonuclease should be inactivated. Phenol/chloroform extraction and ethanol precipitation is an irreversible method for inactivation and removal of all restriction endonucleases...
Added: Tue May 14 2002, Hits: 157, Reviews: 0