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AMPD1 Sigma-Aldrich

DNase I

Amplification Grade

Synonym: Deoxyribonuclease I

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Description

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.

Deoxyribonuclease I (DNase I) is an endonuclease isolated from bovine pancreas that 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.

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.

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.

Safety & Documentation

Safety Information

RIDADR 
NONH for all modes of transport

Frequently Asked Questions

Which document(s) contains shelf-life or expiration date information for a given product?
If available for a given product, the recommended re-test date or the expiration date can be found on the Certificate of Analysis.
How do I get lot-specific information or a Certificate of Analysis?
The lot specific COA document can be found by entering the lot number above under the "Documents" section.
What is the concentration of the DNAse in Product AMPD1, DNase I?
The DNAse is provided at 1 unit/uL in 1 mL total volume.
What do I do if my DNA is not completely digested after 15 minute digestion at room temperature when using Product AMPD1, DNase I?
If further digestion is required, incubation can be performed for 30 minutes at 37°C.
How do I find price and availability?
There are several ways to find pricing and availability for our products.Once you log onto our website, you will find the price and availability displayed on the product detail page.You can contact any of our Customer Sales and Service offices to receive a quote. USA customers: 1-800-325-3010 orview local office numbers.
What is the Department of Transportation shipping information for this product?
Transportation information can be found in Section 14 of the product's (M)SDS.To access the shipping information for this material, use the link on the product detail page for the product.
My question is not addressed here, how can I contact Technical Service for assistance?
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Protocols & Articles

Articles

Transplex® RNA Amplification Kit: Whole Transcriptome Amplification of Highly-Degraded RNA from FFPE Tissues.

The TransPlex Complete Whole Transcriptome Amplification (WTA2 ) Kit effectively amplifies intact and highly degraded RNA. To benchmark maintenance of representative RNA levels during amplification, ...
Ken Heuermann and Brian Ward
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Protocols

Reverse Transcription Using ReadyScript cDNA Synthesis RDRT Protocol

Place components on ice. Mix, and then briefly centrifuge to collect contents at the bottom of the tube
Keywords: Amplification, Digestions, Gene expression, Polymerase chain reaction, Polymerase chain reaction - quantitative, Precipitation, Purification, RNA purification, Transcription

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. Microgram quantit...
Keywords: Amplification, Centrifugation, Condensations, Degradations, Electrophoresis, Eliminations, Enzyme activity, Gene expression, Genomics, Microarray Analysis, Molecular probes, Nucleic acid annealing, Nucleic acid denaturation, Peptide synthesis, Polymerase chain reaction, Polymerase chain reaction - quantitative, Polymerization reactions, Purification, Sample preparations, Sequencing, Sonication

TransPlex® Whole Transcriptome Amplification Protocol

TransPlex, a Whole Transcriptome Amplification (WTA) method, allows for representative amplification of nanogram quantities of total RNA in less than 4 hours without 3'-bias. Microgram quantities of ...
Keywords: Amplification, Centrifugation, Cloning, Condensations, DNA purification, Gene expression, Genomics, Microarray Analysis, Molecular biology, Molecular probes, Nucleic acid annealing, Nucleic acid hybridization, Polymerase chain reaction, Polymerase chain reaction - quantitative, Polymerization reactions, Purification, Sequencing

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Peer-Reviewed Papers
15

References

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