DNase使⽤说明promega
Product Contents
RQ1 RNase-Free DNase:degrade
Part No.Size (units)
M610A1,000
D e s c r i p t i o n:RQ1 (RNA-Qualified) RNase-Free DNase is a DNase I (endonuclease) that degrades both double-stranded and single-stranded DNA, producing 3′-OH oligonucleotides (1). (RQ1 RNase-Free DNase may be used in applications where maintaining the integrity of the RNA is critical.) This DNase is suited for applications such as nick translation (2), production of random fragments (3), cleavage of genomic DNA for footprinting (3), removal of DNA template after in vitro transcription (4), and removal of DNA from RNA samples prior to applications such as RT-PCR (5).
In the presence of Mg2+, DNase I attacks each strand of DNA independently, and the sites of cleavage are distributed in a statistically random fashion (6). In the presence of Mn2+, DNase I cleaves both strands of DNA at approximately the same site to yield fragments with blunt ends or protruding termini
of one or two nucleotides in length (6).
10X Reaction Buffer (M198A): The RQ1 DNase 10X Reaction Buffer provided with this enzyme has a composition of
400mM Tris-HCl (pH 8.0), 100mM MgSO
4and 10mM CaCl
2
.
Enzyme Storage Buffer: RQ1 DNase is supplied in 10mM HEPES (pH 7.5), 50% glycerol (v/v), 10mM CaCl
2
and
10mM MgCl
2
.
Heat Inactivation:10 minutes at 65°C in the presence of Stop Solution.
Inhibitors:EGTA; EDTA (7); salt concentrations >100mM will reduce DNase activity.
Molecular Weight: 31,000 Daltons.
Requirement: Ca2+and Mg2+or Mn2+(7).
Source: Bovine pancreas.
Storage Temperature: Store at –20°C. Avoid exposure to frequent temperature changes. See the expiration date on the Product Information Label.
Stop Solution (M199A): 20mM EGTA (pH 8.0).
Unit Definition:One unit of RQ1 RNase-Free DNase is defined as the amount required to completely degrade 1µg of lambda DNA in 10 minutes at 37°C in 50µl of a buffer containing 40mM Tris-HCl (pH
7.9), 10mM NaCl, 6mM MgCl
2
and 10mM
CaCl
2
. Under these assay conditions one unit of RQ1 DNase activity is approximately equal to one Kunitz unit. See the unit concentration on the Product Information Label.
Usage Notes:
1.This DNase solution does not contain an RNase inhibitor. Observe caution in handling the product to ensure against contaminating it with RNase.
2.Under different buffer conditions the amount of DNase required to completely digest a given amount of DNA may need to be
empirically determined. For example, salt concentrations >100mM will reduce DNase activity.
Quality Control Assays
Contaminant Activity
RNase Assay: To test for RNase activity,50ng of [3H]RNA is incubated with 5 units of RQ1 RNase-Free DNase in Transcription Optimized 1X Buffer (Cat.#P1181, diluted fivefold) for 1 hour at 37°C, and the release of radiolabeled nucleotides is monitored by scintillation counting of TCA-soluble material. The minimum passing specification is <3% release.
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Madison, WI 53711-5399USA
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Toll Free800-356-9526
Fax608-277-2516
Internet
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PRODUCT USE LIMITATIONS, WARRANTY,DISCLAIMER Promega manufactures products for a number of
intended uses. Please refer to the product label for
the intended use statements for specific products. Promega products contain chemicals which may be harmful if misused. Due care should be exercised
with all Promega products to prevent direct human contact.
Each Promega product is shipped with documentation stating specifications and other technical infor
mation. Promega products are warranted to meet or exceed the stated specifications. Promega's sole obligation and the customer's sole remedy is limited to replace-ment of products free of charge in the event products fail to perform as warranted. Promega makes no other warranty of any kind whatsoever, and SPECIFICALLY DISCLAIMS AND EXCLUDES ALL OTHER WAR-RANTIES OF ANY KIND OR NATURE WHATSOEVER, DIRECTLY OR INDIRECTLY, EXPRESS OR IMPLIED, INCLUDING, WITHOUT LIMITATION, AS TO THE SUITABILITY, PRODUCTIVITY, DURABILITY, FITNESS FOR A PARTICULAR PURPOSE OR USE, MER-CHANTABILITY, CONDITION, OR ANY OTHER MAT-TER WITH RESPECT TO PROMEGA PRODUCTS. In
no event shall Promega be liable for claims for any other damages, whether direct, incidental, foresee-able, consequential, or special (including but not lim-ited to loss of use, revenue or profit), whether based upon warranty, contract, tort (including negligence) or strict liability arising in connection with the sale or the failure of Promega products to perform in accordance with the stated specifications.
Part# 9PIM610
Revised 2/05
Part# 9PIM610
Printed in USA. Revised 2/05
Riboprobe is a trademark of Promega Corporation and is registered with the U.S. Patent and Trademark Office.? 1996–2005 Promega Corporation. All Rights Reserved.
All specifications are subject to change without prior notice.
Product claims are subject to change. Please contact Promega Technical Services or access the Promega online catalog for the most up-to-date information on Promega products.
AF9PI M6100205M610
(a)The PCR process is covered by patents issued and applicable in certain countries. Promega does not encourage or support the unauthorized or unlicensed use of the PCR process. Use of this product is recommended for persons that either have a license to perform PCR or are not required to obtain a license.
(b)U.S. Pat. No. 5,552,302, European Pat. No. 0 422 217, Australian Pat. No. 646803 and Japanese
Pat. Nos. 3009458 and 3366596 have been issued to Promega Corporation for the methods and compositions for production of human recombinant placental ribonu-clease inhibitor.
(c)U.S. Pat. Nos. 4,966,964, 5,019,556 and 5,266,687, Australian Pat. Nos. 616881 and 641261 and other pending and issued patents, which claim vectors encoding a portion of human placental ribonuclease inhibitor, are exclusively licensed to Promega Corporation.
Usage Information

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