T4 RNA Ligase
Joins ssRNA or ssDNA onto the 5´-Phosphoryl End of ssRNA or ssDNA
- Adds [5´-32P] nucleoside 3´,5´-bis (phosphate) onto single-stranded RNA
- Can be inactivated by heating at 65°C for 15 minutes
- Supplied at a concentration of 10u/μl
Catalog Number:
Size
Catalog Number: M1051
T4 RNA Ligaseは、ATPに依存して、1本鎖RNAまたはDNAの分子間および分子内の連結を行い、1本鎖RNAまたはDNAの5'-末端のリン酸基と3'-末端の水酸基を結合します。同様に、1本鎖RNAに[5'-32P]nucleoside3',5'-bis(phosphate)を付加する反応も行います。本酵素はRNase Iを欠如する組換えE.coli CA4株から精製しており、分子量は約43.5KDaです。
- 10X Reaction Buffer添付： 500mM Tris-HCl(pH7.8 at25℃),100mM MgCl2,50mM DTT,10mM ATP
- 熱失活： 65℃,15分で不活性化します
- 1本鎖RNAまたはDNAの連結
- RNAの3′-末端標識
- RNAおよびDNAオリゴヌクレオチドの環状化
- 完全長のcDNAのクローニング
- Silber, R. et al. (1972) Proc. Natl. Acad. Sci. USA 69, 3009–13.
- England, T. et al. (1977) Proc. Natl. Acad. Sci. USA 74, 4839–42.
- Uhlenbeck, O.C. and Gumport, R.I. (1982) The Enzymes 158, 31.
- Brennan, C.A. et al. (1983) Meth. Enzymol. 100, 38–52.
- Schaefer, B.C. (1995) Anal. Biochem. 227, 255–73.
Storage Buffer: 10mM Tris (pH 7.5 at 25°C), 50mM KCl, 0.1mM EDTA, 1mM DTT, 50% glycerol, 0.1% Tween® 20.
Source: Recombinant E. coli strain.
QC Tests: Activity, SDS-PAGE/purity, DNase, RNase, endonuclease.
Unit Definition: One unit catalyzes the formation of 1nmol of [5´-32P]rA14–20 into a phosphatase-resistant form in 30 minutes at 37°C in a total volume of 30μl. The reaction conditions are: 50mM Tris-HCl (pH 7.8 at 25°C), 10mM MgCl2, 5mM DTT, 1mM ATP and 10μM [5´-32P]rA14–20.
Protocols
Complete Protocol
Specifications
Catalog Number:
選択製品の構成品内容
| Item | Part # | Size |
|---|---|---|
|
T4 RNA Ligase |
M105A | 1 × 500u |
|
T4 RNA Ligase 10X Buffer |
M107A | 1 × 500μl |
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