DNase I functions by hydrolyzing phosphodiester linkages, producing mono and oligonucleotides with 5'-phosphate and 3'-hydroxyl group. RNase-free DNase I is recommended to degrade DNA in presence of RNA, in absence of RNase is critical to maintain integrity of RNA. For example, DNase I is frequently used to remove template DNA following in vitro transcription, and to remove contaminating DNA in total RNA preparations (especially those from transfected cells that may contain plasmid DNA), used for ribonuclease protection assays, cDNA library contraction, and RT-PCR.
Tested for contaminating RNase and protease activity
Functionality is determined by digestion of human genomic DNA followed by quantitative real-time PCR to detect undigested DNA
Requires bivalent cations (Mg2+ and Ca2+ at approximately 5mM and 0.5mM, respectively) for maximal activity, and has a pH optimum of 7.8
Unit definition: One unit is amount of enzyme required to completely degrade 1μg DNA in 10 min. at 37°C, and is equivalent to 0.04 Kunitz units
For an alternative to bovine DNase I, please consider Recombinant DNase I (Mfr. No. AM2235). For a more active, salt-tolerant DNase, please see the TURBO™ DNase products (Mfr. Nos. AM2239 and AM2238).
PCR & Real-Time PCR, Real Time PCR (qPCR), Reverse Transcription