T7428
Topoisomerase I from wheat germ
buffered aqueous glycerol solution
Synonym(s):
DNA relaxing enzyme, Topo I
grade
Molecular Biology
form
buffered aqueous glycerol solution
mol wt
97 kDa
foreign activity
RNase and DNase, none detected
shipped in
dry ice
storage temp.
−70°C
General description
Topoisomerase I relaxes supercoiled DNA molecules. The enzyme initiates transient breakages and rejoins of phosphodiester bonds in superhelical turns of closedcircular dsDNA.
Application
Suitable for:
- Analsis of degree of DNA supercoiling
- Forming knots and circles in single stranded DNA
- Converting complementary single stranded DNA to double stranded, circular forms
- Rejoining nicks in double-stranded DNA
Biochem/physiol Actions
Topoisomerase I relaxes supercoiled DNA molecules. The enzyme initiates transient breakages and rejoins of phosphodiester bonds in superhelical turns of closed-circular DNA. Enzyme activity is independent of right- and left-handed superhelices.
Other Notes
One unit will convert 1μg of supercoiled pGEM9Zf(−) plasmid DNA to the relaxed form in 30 min at 37 °C.
To preserve enzyme activity, avoid multiple freeze-thaw cycles and frequent temperature variations.
Topoisomerase I is supplied in a solution of 50 mM Tris-HCl, pH 7.9, 1 mM EDTA, 1 mM DTT, 0.5 M NaCl and 50% glycerol (v/v).
Storage Class
11 - Combustible Solids
wgk
WGK 3
flash_point_f
Not applicable
flash_point_c
Not applicable
ppe
Eyeshields, Gloves, multi-purpose combination respirator cartridge (US)
Regulatory Information
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P D Foglesong et al.
BioTechniques, 13(3), 402-404 (1992-09-01)
Supercoiled and relaxed DNA were resolved electrophoretically in the presence of 0.5 micrograms/ml ethidium bromide. Under these conditions the Gaussian distributions of topological isomers of both supercoiled and relaxed DNA migrated as discrete bands. The separation of these DNAs was
J C Wang
Annual review of biochemistry, 65, 635-692 (1996-01-01)
The various problems of disentangling DNA strands or duplexes in a cell are all rooted in the double-helical structure of DNA. Three distinct subfamilies of enzymes, known as the DNA topoisomerases, have evolved to solve these problems. This review focuses
Maris A Cinelli et al.
Journal of medicinal chemistry, 55(24), 10844-10862 (2012-12-12)
Hydroxylated analogues of the anticancer topoisomerase I (Top1) inhibitors indotecan (LMP400) and indimitecan (LMP776) have been prepared because (1) a variety of potent Top1 poisons are known that contain strategically placed hydroxyl groups, which provides a clear rationale for incorporating
Tomoko Nanbu et al.
Molecular and cellular biology, 33(6), 1175-1187 (2013-01-09)
Protection of telomeres protein 1 (Pot1) binds to single-stranded telomere overhangs and protects chromosome ends. RecQ helicases regulate homologous recombination at multiple stages, including resection, strand displacement, and resolution. Fission yeast pot1 and RecQ helicase rqh1 double mutants are synthetically
Xiaoduo Zhi et al.
Gene, 514(2), 82-90 (2012-12-04)
Transcription by RNA polymerase can induce the formation of hypernegatively supercoiled DNA in vitro and in vivo. This phenomenon has been nicely explained by a "twin-supercoiled-domain" model of transcription where a positively supercoiled domain is generated ahead of the RNA
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