商品编号


EG-150



品牌


Excellgen



公司


Excellgen, Inc.



公司分类


Polymerases




1,000 Units

商品信息



Description


Yeast Poly(A) Polymerase uses ATP as a substrate for template-independent addition of AMP from ATP to the 3? hydroxyl termini of RNA molecules. It can be used to add poly(A) tails to RNA in the first step of cloning. The reaction requires Mn2+ or Mg2+, ATP as substrate, and any RNA containing 3' hydroxyl termini as primer. Substitution of cordycepin-5'-triphosphate (3'-dATP) for ATP results in addition of a single 3'-dA residue to the ends of the RNA, a useful technique for labeling RNA at the 3' end.

In comparative studies, Yeast Poly(A) Polymerase works more efficiently than E. coli poly(A) polymerase for RNA oligonucleotide-labeling and poly(A) tailing. Shorter incubation times are required for the yeast enzyme and it is found to label both long and short substrates equally well. Yeast Poly(A) Polymerase is recommended over T4 RNA Ligase for 3'-end labeling of long RNA molecules.




Applications




Addition of a poly(A) tail to RNA synthesized in vitro for increasing RNA st
ABI
lity and translation efficiency prior to transferring into eukaryotic cells



CDN
A library construction using a primer containing poly(dT) on its 3? end.



Synthesis of polyadenylated RNA for nucleic acid amplification






Source

A recombinant
E. coli
strain carrying yeast
(Saccharomyces cerevisiae)
Poly(A) Polymerase gene



Unit Definition

1 unit is defined as the amount of enzyme that will incorporate 1 nmol of ATP into acid-insoluble material in 10 minutes at 37°C.



Components



Yeast Poly(A) Polymerase:
5,000 U/ml in 20 mM Tris-HCl, 300 mM NaCl, 1 mM DTT, 1 mM EDTA, 50% Glycerol, 0.1% Triton? X-100, pH 7.5 @ 25°C

5X Reaction Buffer
: 1500 mM Tris-HCl, pH 7.0, 3.0 mM MnCl2, 0.1 mM EDTA, 1 mM DTT, 500 μg/ml acetylated BSA, 50% glycerol.





Quality Control

The absence of endonuclease, nicking activity,

exonuclease and RNAse

activity is confirmed by appropriate quality tests.



Storage Condition


-20 °C