Thermo Fisher Scientific Ribonuclease H, E.coli User guide

Brand: Thermo Fisher Scientific

Model: Ribonuclease H, E.coli

Type: User guide

Ribonuclease H, E.coli (Cloned)

Catalog Number AM2292, AM2293

Pub. No. MAN0019139 Rev. A.00

Contents and storage

Cat. No.

Contents

Source

Amount

Storage

AM2292

Ambion® Ribonuclease

H, E.coli

An E.coli strain

overexpressing RNase H.

200 Units, 10 U/µL

Store at –25 °C to -15 °C. Do not

store in a frost-free freezer.

AM2293

1000 Units, 10 U/µL

Product description

A high purity Ribonuclease H, suitable for most RNA amplification and nucleic acid sequence-based amplification (NASBA)

protocols.

Unit definition

One unit of the enzyme catalyzes the formation of 1 nmol of acid soluble products in 20 min at 37 °C.

Storage buffer

20 mM Tris-HCl (pH 7.5 at 25 °C), 0.1 mM EDTA, 100 mM KCl, 10 mM MgCl2, 1 mM DTT, 0.02 % Triton X100, 50 % Glycerol.

General information

Ribonuclease H (RNase H) specifically degrades the RNA in RNA:DNA hybrids to produce 3'−hydroxyl and 5'−phosphate

terminated products. The enzyme will not degrade single-stranded DNA, double-stranded DNA, or unhybridized RNA. RNase H

is an essential component of the Ambion MessageAmp™ II aRNA Amplification Kit (P/N AM1751), which is based on the

patented Eberwine aRNA amplification procedure. RNase H can also be used to degrade a specific RNA when the

complementary DNA oligonucleotide is hybridized to it, as is used in antisense technology, and for poly(A) tail removal from

mRNA hybridized to oligo(dT).

Applications

The following general protocol for digestion of RNA;DNA hybrids is adapted from Current Protocols in Molecular Biology. The

general reaction parameters, including the amount of enzyme, and the method of stopping the reaction, will vary depending on

your experimental needs.

Digestion of RNA:DNA hybrids

20 mM HEPES-KOH, pH 8.0

50 mM KCl

4 mM MgCl2

1 mM DTT

2 μg RNA:DNA hybrid

50 μg/mL BSA

~10 unit Ribonuclease H per μg RNA:DNA hybrid

Total reaction volume: 100 μL

Incubate 20 min at 37 °C

Add 1 μL 0.5 M EDTA to stop the reaction.

For Research Use Only. Not for use in diagnostic procedures.

WARNING! Read the Safety Data Sheets (SDSs) and follow the handling instructions. Wear appropriate protective eyewear, clothing, and gloves. Safety

Data Sheets (SDSs) are available from thermofisher.com/support.

Reference

Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Struhl K, editors. (2006) Enzymatic

Manipulation of DNA and RNA (Chapter 3). In Current Protocols in Molecular Biology. John Wiley & Sons, Inc.

p 3.13.2.

Limited product warranty

Life Technologies Corporation and/or it affiliate(s) warrant their products as set forth in the Life Technologies’ General Terms and Conditions

of Sale at www.thermofisher.com/us/en/home/global/terms-and-conditions.html. If you have any questions, please contact Life

Technologies at www.thermofisher.com/support.

The information in this guide is subject to change without notice.

DISCLAIMER: TO THE EXTENT ALLOWED BY LAW, THERMO FISHER SCIENTIFIC INC. AND/OR ITS AFFILIATE(S) WILL NOT BE LIABLE FOR SPECIAL, INCIDENTAL, INDIRECT,

PUNITIVE, MULTIPLE, OR CONSEQUENTIAL DAMAGES IN CONNECTION WITH OR ARISING FROM THIS DOCUMENT, INCLUDING YOUR USE OF IT.

Important Licensing Information: These products may be covered by one or more Limited Use Label Licenses. By use of this product, you accept the terms and conditions of all

applicable Limited Use Label Licenses.

hermofisher.com/support | thermofisher.com/askaquestion

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Thermo Fisher Scientific Baltics UAB | V.A. Graiciuno 8, LT-02241 Vilnius, Lithuania

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26 February 2020

Thermo Fisher Scientific Ribonuclease H, E.coli User guide

Brand: Thermo Fisher Scientific

Model: Ribonuclease H, E.coli

Type: User guide

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Thermo Fisher Scientific Ribonuclease H, E.coli User guide

Thermo Fisher Scientific Ribonuclease H, E.coli User guide

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