Irreversible Heat Inactivation of DNase I without RNA Degradation

Irreversible heat inactivation of DNase I without RNA degradation.

In many applications of RT-PCR, residual genomic DNA in total RNA preparations is amplified together with RNA, which results in false-positive data. One way to overcome this problem is to design primers that span a region containing one or more introns, thereby creating a diagnostic size difference between the amplification products originating from the RNA or the DNA. However, this method will...

DNase I activity retained after heat inactivation in standard buffer.

For the detection of RNA transcripts by RT-PCR, prior removal of genomic DNA must be performed. To remove genomic DNA, RNA is often prepared by DNase I digestion following phenol extraction. Recently, one-tube or onebuffer systems of RT-PCR were developed to prevent loss of RNA and to reduce the risk of contamination (2,6). In these methods, DNase I is added before RT. Taq DNA polymerase prepar...

DNaseR: DNase I footprinting analysis of DNase-seq data

The combination of DNase I digestion and high-throughput sequencing (DNaseseq) has been used recently to map chromatin accessibility in a given tissue or cell type on a genome-wide scale (Song and Crawford, 2010). In addition to DNase I hypersensitive sites (DHSs), short regions of protected nucleotides known as footprints can be detected using a technique known as ”digital genomic footprinting...

DNase I treatment of total RNA improves the accuracy of ribonuclease protection assay.

DNase I digestion of chromatin from avian liver nuclei liberates DNA-dependent RNA polymerase II.

In an effort to develop mild conditions for the isolation of DNA-dependent RNA polymerase II, we have used DNase I covalently coupled to Sepharose 4B to digest chromatin from hypotonically lysed nuclei from rooster liver. The RNA polymerase II released was at least 2 times more active in in vitro transcription than was RNA polymerase II prepared by the conventional method of sonication of chrom...