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Protocols
- 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
DNA polymerase
http://www.oup.co.uk/pdf/pas/9v1-7-10.pdf
Added: Sat Aug 17 2002, Hits: 2244, Reviews: 0
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Applications of Modifying Enzymes
(NEB)
Presents quick reference table for application of modifying enzymes.
http://www.neb.com/neb/tech/tech_resource/modifyin...
Added: Tue May 14 2002, Hits: 81, Reviews: 0
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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.
http://hg.wustl.edu/hdk_lab_manual/rdm/rdm9.html
Added: Tue May 14 2002, Hits: 380, Reviews: 0
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Fill-In Reaction
(Hedrick Lab, UCSD Cancer Center)
Reaction for generating blunt ends on dsDNA fragments with overhangs (especially good for 5' overhangs).
http://hedricklab.ucsd.edu/Protocol/FillIn.html
Added: Tue Dec 10 2002, Hits: 584, Reviews: 0
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Thermal Inactivation
(Fermentas)
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...
http://www.fermentas.com/techinfo/RE/restrthermina...
Added: Tue May 14 2002, Hits: 158, Reviews: 0
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