226 Maunders
The temperature of reaction must be optimized for the specific appli-
cation, conditions ranging from 37°C for 30 min to 4°C overnight.
After this time, the reaction should be terminated by boiling for 2 min.
- Summary
DNA and RNA ligases are widely occurring, readily available, and
relatively inexpensive enzymes that are an essential part of the molecular
biologist's armory, both for the analysis and the manipulation of nucleic
acids. The major drawback of these enzymes lies in their relative lack
of substrate specificity as far as nucleotide sequence is involved. This
factor necessitates careful experimental design in order to obviate the
production of spurious or misinterpreted results.
References
- Engler, M. J. and Richardson C. C. (1982) DNA ligases, in The Enzymes XV
Part B (Boyer, P. D., ed.), Academic, New York, pp. 3-29.
- Gellert, M. (1967) Formation of covalent circles of lambda DNA by E. coli
extracts. Proc. Natl. Acad. Sci. USA 57, 148-155.
- Laipis, P. J., Olivera, B. M., and Ganesan, A.T. (1969) Enzymatic cleavage
and repair of transforming DNA. Proc. Natl. Acad. Sci. USA 62, 289-296.
- Weiss, B. and Richardson, C. C. (1967) Enzymatic breakage and joining of
deoxyribonucleic acid. I. Repair of single strand breaks in DNA by an enzyme
system from Escherichia coli infected with T4 bacteriophage. Proc. Natl. Acad.
Sci. USA 57, 1021-1028.
- Soderhall, S. and Lindhahl, T. (1976) DNA ligases of eukaryotes. FEBS Lett.
67, 1-7.
- Pfeiffer, P. and Vielmetter, W. (1988) Joining of nonhomologous DNA double
strand breaks in vitro. Nucleic Acids Res. 16, 907-924.
- North, P., Ganesh, A., and Thacker, J. (1990) The joining of double strand
breaks in DNA by human cell extracts. Nucleic Acids Res. 18, 6205-6210.
- Okasaki, K., Okasaki, T., Sakabe, K., Sugimoto, K., and Sugino, A. (1968) Mech-
anism of DNA chain growth. I. Possible discontinuity and unusual secondary
structure of newly synthesized chains. Proc. Natl. Acad. Sci. USA 59, 598-605.
- Boyce, R. P. and Howard-Flanders, P. (1964) Release of ultraviolet light induced
thymine dimers from DNA of E. coli K-12. Proc. Natl. Acad. Sci. USA 51,
293-300.
- Johnston, L. H. and Nasmyth, K. A. (1978) Saccharomyces cerevisiae cell
cycle mutant cdc9 is defective in DNA ligase. Nature (London) 274, 891-893.
- Panasenko, S. M., Cameron, J. R., Davis, R. W., and Lehman, I. R. (1977)
Five hundred-fold overproduction of DNA ligase after induction of a hybrid
lambda lysogen constructed in vitro. Science 196, 188-189.
- Panasenko, S. M., Alazard, R. J., and Lehman, I. R. (1978) A simple three step
procedure for the large scale purification ofDNA ligase from a hybrid lambda
lysogen constructed in vitro. J. Biol. Chem. 253, 4590-4592.