User:Jarle Pahr/Enzymes: Difference between revisions
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Klenow fragment: Fragment of E. coli DNA polymerase I, retaining 5'-3' polymerase activity and 3’ → 5’ exonuclease activity, but without 5' → 3' exonuclease activity. The 3'->5' exonuclease activity of the Klenow fragment is roughly 200 times less than that of T4 DNA polymerase (http://www.vivo.colostate.edu/hbooks/genetics/biotech/enzymes/t4dnap.html). Klenow fragment is commonly used to generate blunt ends by overhang fill-in. | Klenow fragment: Fragment of E. coli DNA polymerase I, retaining 5'-3' polymerase activity and 3’ → 5’ exonuclease activity, but without 5' → 3' exonuclease activity. The 3'->5' exonuclease activity of the Klenow fragment is roughly 200 times less than that of T4 DNA polymerase (http://www.vivo.colostate.edu/hbooks/genetics/biotech/enzymes/t4dnap.html). Klenow fragment is commonly used to [http://www.mdanderson.org/education-and-research/departments-programs-and-labs/labs/dent-laboratory/protocols/klenow-t4-dna-polymerase-end-blunting.html generate blunt ends] by overhang fill-in. | ||
See also the book [http://link.springer.com/book/10.1385/0896032345/page/1 Enzymes of molecular biology] | See also the book [http://link.springer.com/book/10.1385/0896032345/page/1 Enzymes of molecular biology] | ||
Revision as of 09:07, 15 March 2013
Notes on various enzymes:
Polymerases (Not used in PCR. For polymerases used in PCR, see PCR page):
T4 DNA polymerase:
Posesses 5' -> 3 DNA polymerase and 3' -> 5' exonuclease activity. Used in SLIC. Does not show strand displacement activity.
T5 Exonuclease: A 5' -'3 exonuclease. Used in Gibson Assembly.
Klenow fragment: Fragment of E. coli DNA polymerase I, retaining 5'-3' polymerase activity and 3’ → 5’ exonuclease activity, but without 5' → 3' exonuclease activity. The 3'->5' exonuclease activity of the Klenow fragment is roughly 200 times less than that of T4 DNA polymerase (http://www.vivo.colostate.edu/hbooks/genetics/biotech/enzymes/t4dnap.html). Klenow fragment is commonly used to generate blunt ends by overhang fill-in.
See also the book Enzymes of molecular biology
