PEG in ligations, why?-References
Bradley Turner
bsturner at MBCRR.HARVARD.EDU
Fri Jun 7 01:35:00 EST 1996
Troels,
The following references discuss some aspects of PEG ligation enhancement:
Polymer-stimulated ligation: enhanced blunt- or cohesive-end ligation
of DNA or deoxyribonucleotides by T4 DNA ligase in polymer solutions
Barbara H. Pheiffer and Steven B. Zimmerman
Nucleic Acids Research 11(22):7853-7871, 1983
Macromolecular crowding allows blunt-end ligation by DNA ligases
from rat liver or Escherichia coli
Steven B. Zimmerman and Barbara H. Pheiffer
Proc Natl Acad Sci USA 80:5852-5856, 1983 Oct
Macromolecular crowding accelerates the cohesion of DNA fragments
with complementary termini
S. B. Zimmerman and B Harrison
Nucleic Acids Research 13:2241
I generally use a final concentration of 5-15%(w/v) of PEG8000 in my ligation
reactions-you can prepare a 40%(w/v) stock solution and dilute it
appropriately.
I hope this helps,
Brad,
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Bradley Turner
Beth Israel Hospital
Harvard Medical School 617-667-1215 phone
Division of Gastroenterology 617-667-2767 fax
Room Dana 536 turner at sprcore.bih.harvard.edu
330 Brookline Avenue bsturner at mbcrr.harvard.edu
Boston, MA 02215 bturner at bih.harvard.edu
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