Description

    Recombinant RNase Inhibitor is expressed in E. coli, containing a plasmid that carries the porcine RNase Inhibitor gene,and purified by affinity chromatography. This product is similar to the one from human placenta, and can be used in the same applications as RNase inhibitors derived from human placenta. This product forms a 1:1 complex with RNase A, and inhibits RNase activity noncompetitively. Because it is a protein, it differs from other competitive inhibitors (nucleotides and inorganic phosphatases) in that it can easily be removed from the reaction system by phenol extraction. However, it does not inhibit the RNase H activity of reverse transcriptase. This Inhibitor can be added directly to reaction mixtures containing RNA for cDNA synthesis，In vitro translation，In vitro transcription and Polysome isolation.

Components

    Recombinant RNase Inhibitor（Porcine)

    RIN-002-4KU                   4KU/100µl

    RIN-002-40KU                  40KU/1ml

    RIN-002-2000KU                2000KU/50ml

STORAGE BUFFER

    20 mM HEPES-KOH(pH 7.6), 50mM KCl, 8mM DTT, 50% (v/v) glycerol. 

Unit definition

    One unit is defined as the amount of Recombinant Ribonuclease Inhibitor required to inhibit the activity of 5 ng of RNase A by 50%.

QUALITY CONTROL

    -Each batch of RNase Inhibitor is confirmed to contain  <2% contaminating protein by SDS-PAGE.                       
    -Each batch of RNase Inhibitor is confirmed to inhibit the activity of 10 ng of RNase A in 1-step RT-qPCR with 25µl reaction volume.  

    -RNase Inhibitor are subjected to stringent quality control tests, are free of contamina- ting exo-and endonuclease activity, and meet strict requirements with respect to DNA contamination levels . 

Notes

    -This inhibitor acts as a noncompetitive inhibitor of RNases A, B and C and is active over a wide pH range and in at least 1 mM DTT.

    -This inhibitor does not inhibit the RNase H activity of reverse transcriptase.

Applications

    -cDNA synthesis (Ribonuclease Inhibitor: 0.5 U/µl reaction)

-1-step RT-qPCR (0.3-0.5 U/µl reaction)

-cDNA synthesis ( 0.3-0.5 U/µl reaction)

    -In vitro translation (1 U/µl reaction)

    -In vitro transcription using cell-free extract (20 U/µl reaction)

    -In vitro transcription with SP6 or T7 RNA Polymerase (1 U/µl reaction)

    -Polysome isolation (1,000 U/ml reaction)

STORAGE

This product should be kept at -20°C for 12 months