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AMPD1

Sigma-Aldrich

DNase I

Amplification Grade

Synonym(s):
Deoxyribonuclease I
Enzyme Commission number:
NACRES:
NA.55

Quality Level

form

liquid

concentration

1 unit/μL

color

colorless

shipped in

wet ice

storage temp.

−20°C

General description

Deoxyribonuclease I (DNase I) is an endonuclease isolated from bovine pancreas that digests double and single stranded DNA into oligo and mononucleotides. Amplification Grade DNase I has been purified to remove RNase activity, and is suitable for eliminating DNA from RNA preparations prior to sensitive applications, such as RT-PCR (Reverse Transcriptase - Polymerase Chain Reaction).

DNase I digests double- and single-stranded DNA into oligo- and mononucleotides. Using the Reaction Buffer provided, DNA is removed from RNA preparations in a 15 minute digestion at room temperature. The DNase I is then inactivated by heating with the Stop Solution. Heating also denatures hairpins in the RNA, so the RNA can be used directly in reverse transcription.

No current RNA isolation procedure removes 100% of the DNA. Many commercial DNase I formulations are contaminated with residual RNases. This RNase contamination can destroy or degrade valuable RNA samples prior to reverse transcription. Laboratory comparisons have shown that Sigma′s Amplification Grade DNase I demonstrates lower RNase activity than that from several leading molecular biology product suppliers.

Application

Amplification grade DNase I has been used:
  • for the digestion of DNA during isolation and purification of RNA. The purified RNA can be used for the synthesis of cDNA using RNA reverse transcriptase.
  • to hydrolyze extracellular matrix (ECM) components and enhance photosensitizer penetration into the biofilm to determine the efficacy of antimicrobial photodynamic therapy (aPDT) on Candida albicans biofilms
  • to remove contaminating DNA from total RNA extracted from cattle blood samples

Features and Benefits

  • Suitable for the elimination of DNA from RNA
  • Minimal RNase activity available
  • Optimized 10× reaction buffer and Stop Solution for complete inactivation of DNase I

Suitability

Suitable for use in removing DNA from RNA preparations.

Unit Definition

One unit completely digests 1 μg of plasmid DNA to oligonucleotides in 10 min. at 37 °C.

Legal Information

Purchase of this product is accompanied by a limited license for use in the Polymerase Chain Reaction (PCR) process for research purposes only and in conjunction with a thermal cycler whose use in the automated performance of the PCR process is covered by an up-front license fee, either by payment to Applied Biosystems or as purchased, i.e., and authorized thermal cycler.

Storage Class Code

10 - Combustible liquids

WGK

WGK 2

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Certificate of Analysis

Enter Lot Number to search for Certificate of Analysis (COA).

Certificate of Origin

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Protocols

Reverse Transcription Protocol

Method for reverse transcription of RNA into DNA. Uses a premixed reagent that contains reverse transcriptase, dNTPs, primers, RNase inhibitor and buffer. Fast generation of cDNA.

SEQR - SeqPlex RNA Amplification Kit Protocol

The SeqPlex RNA Amplification kit provides a method for amplification of total RNA or isolated mRNA prior to entry into the workflows of the commonly used deep sequencing platforms.

Related Content

Spectrum™ Plant Total RNA Kit

The Spectrum Plant Total RNA Kit incorporates a new, patent-pending lysis and binding chemistry. This kit is intended to purify total RNA from species and tissue where most Total RNA protocols fail. Without sacrificing affordability, you can purify more total RNA per isolation.

PCR Applications

Polymerase chain reaction (PCR) is a technique for amplifying nucleic acid molecules and is commonly used in many applications, including RT-PCR, hot start PCR, end point PCR and more.

PCR Applications

Polymerase chain reaction (PCR) is a technique for amplifying nucleic acid molecules and is commonly used in many applications, including RT-PCR, hot start PCR, end point PCR and more.

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