M0463S, yDcpS - 4000 units

yDcpS is the scavenging decapping enzyme from S. cerevisiae that hydrolyzes the phosphodiester bond between the gamma and beta phosphates of m⁷G-capped mRNA, producing diphosphorylated 5′ ends and m⁷GMP.yDcpS is capable of decapping mRNAs of various lengths, decaps Cap0 and Cap1 with equal efficiency, and the diphosphorylated 5′ end it produces is suitable for recapping with Vaccinia Capping Enzyme.

Product Source

A His-tagged engineered variant of E. coli that carries the S. cerevisiae gene DCS1 (encoding yDcpS) on a plasmid.

Unit Definition

One unit is defined as the amount of yDcpS required to convert 50% of a 500 nM solution of the following 25-mer m7G-capped RNA to a 5’-diphosphorylated form in a total reaction volume of 20 µl in 1 hour at 37˚C: 5’-[m7Gppp]rGrUrArGrArArCrUrUrCrGrUrCrGrArGrUrArCrGrCrUrCrArA[3-6FAM]-3’

Reaction Conditions

1X yDcpS Reaction Buffer
Incubate at 37°C

1X yDcpS Reaction Buffer
10 mM Bis-Tris-HCl
1 mM EDTA
(pH 6.5 @ 25°C)

Storage Buffer

300 mM NaCl
10 mM Tris-HCl
0.1 mM EDTA
1 mM DTT
50% Glycerol
pH 7.4 @ 25°C

Heat Inactivation

70°C

Specific Activity

20,000 units/mg

Unit Assay Conditions

1X yDcpS Reaction Buffer with 500 nM m⁷G-capped 25-mer 3´-FAM RNA. After incubation at 37°C for 1 hour, yDcpS is inactivated at 70˚C for 15 minutes and decapping is measured by capillary electrophoresis.