Description
General description
The On-Column DNase I Digestion Set offers a simple and convenient method for eliminating genomic DNA from total RNA preparations. This Set is intended for use with the GenElute™ Mammalian Total RNA Miniprep Kit and the Spectrum Plant Total RNA Kit. It can also be used with many other commercially available RNA purification kits.
The vast majority of DNA is eliminated from total RNA purifications by the procedures provided in the standard protocols of the Sigma Kits named below. However, for very sensitive applications, such as quantitative RT-PCR, even trace amounts of DNA can result in false positive detection of cDNA-derived amplicons.
The On-Column DNase I Digestion Set offers a simple and convenient method for eliminating genomic DNA from total RNA preparations. This Set is intended for use with the GenElute™ Mammalian Total RNA Miniprep Kit, the GenElute Bacterial Total RNA Purification Kit, the GenElute Yeast Total RNA Purification Kit and the Spectrum™ Plant Total RNA Kit. However, the digestion protocol is compatible with many other commercially available RNA purification kits.
The term ′On-Column′ indicates that digestion is accomplished in-process for true convenience. Each of Sigma′s total RNA purification kits utilize a binding column for purification. DNase I digestion is accomplished in 15 minutes while total RNA is bound to the column substrate. Several wash steps and elution of the total RNA immediately follow. To review the complete protocol, see the Technical Bulletin for any of the Kits named above.
Application
On-Column DNase I Digestion Set is suitable for removing gDNA from total RNA preparations. It may not be suitable for RT-PCR applications as a small amount of residual DNA may remain. On-Column DNase I Digestion Set was used to purify RNA preparations isolated from Vitis vinifera cultivars.
Biochem/physiol Actions
The on-column digestion is accomplished in-process for convenience. DNase I digestion is accomplished in 15 minutes while total RNA is bound to the column substrate. Several wash steps and elution of the total RNA immediately follow.
Features and Benefits
The On-Column DNase I Digestion Set offers a simple and convenient method for eliminating genomic DNA from total RNA preparations. This Set is intended for use with the GenElute™ Mammalian Total RNA Miniprep Kit, the GenElute Bacterial Total RNA Purification Kit, the GenElute Yeast Total RNA Purification Kit and the Spectrum™ Plant Total RNA Kit. However, the digestion protocol is compatible with many other commercially available RNA purification kits.
Legal Information
GenElute is a trademark of Sigma-Aldrich Co. LLC
Spectrum is a trademark of Sigma-Aldrich Co. LLC
