DNase I, Amplification Grade

Synonyms: Deoxyribonuclease I

Storage: -20C

Enzyme Commission (EC) Number: 3.1.21.1  ( BRENDA | IUBMB )

UNSPSC Code: 12352200

Features and Benefits: . Suitable for the elimination of DNA from RNA . Minimal RNase activity available. Optimized 10x reaction buffer and Stop Solution for complete inactivation of DNase I

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: 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.

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.

Suitability: Suitable for use in removing DNA from RNA preparations.

Unit Definition: One unit completely digests 1 mug of plasmid DNA to oligonucleotides in 10 min. at 37 C.