DNase I Treatment Kit

Synonyms

RNA, DNase, sample preparation, DNA, lncRNA, mRNA, miRNA, RNA-seq

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Overview

The DNase I Treatment Kit utilizes an endonuclease that non-specifically cleaves ssDNA, dsDNA and the DNA strand of RNA:DNA hybrids to improve the purity of your RNA samples. Following DNase I treatment, RNA samples are purified with easy-to-use spin columns. RNA is suitable for use in downstream applications such as RT-qPCR and RNA-seq.

Description

Kit Specifications:

• Efficiently remove genomic DNA from RNA samples
• Cleaves ssDNA, dsDNA and the DNA strand of RNA:DNA hybrids
• Includes RNA purification reagents for cleanup following DNase I treatment

Advantages of DNase I Treatment:

• Eliminate any trace genomic DNA contamination from your RNA sample
• Avoid reverse transcription quantitative PCR (RT-qPCR) bias that could be caused by contaminating genomic DNA serving as the PCR template
• Increase the accuracy of your RNA-seq by removing all the DNA from the sample to ensure the sequencing reads are specific to the RNA sample

Figure 1: Agarose gel of nuclear and cytoplasmic RNA fractions.

RNA was isolated as nuclear and cytoplasmic fractions using the RNA Subcellular Isolation Kit (Active Motif Catalog No. 25501). Following purification, half of the nuclear RNA was treated with DNase I to remove genomic DNA contamination using the DNase I Treatment Kit. 1 µg of each RNA fraction was loaded onto a 1.5% gel for analysis.
Lane M = 100 bp DNA ladder
Lanes 1-2 = Cytoplasmic RNA
Lanes 3-4 = Nuclear RNA before DNase I treatment
Lanes 5-6 = Nuclear RNA after DNase I treatment

Figure 2: TapeStation analysis of nuclear RNA fractions before and after DNase I treatment.

Nuclear RNA was isolated using the RNA Subcellular Isolation Kit (Active Motif Catalog No. 25501). Following purification, half of the nuclear RNA was treated with DNase I to remove genomic DNA contamination using the DNase I Treatment Kit. 1 µg of each RNA fraction was loaded onto a DNA tape for analysis. The low MW bands represent the DNA marker for each sample.
A1, B1 = Nuclear RNA before DNase I treatment
C1, D1 = Nuclear RNA after DNase I treatment

Contents

Contents & Storage

Please note that the DNase I Treatment Kits are shipped on dry ice and contain reagents with multiple storage temperatures inside. Please store each component at the temperature indicated below. All reagents are guaranteed stable for 6 months from date of receipt when stored properly.This kit includes the following components:

• DNase I (2 U/µl); Store at -20°C
• DNase I Stop Solution; Store at 4°C
• 10X DNase I Reaction Buffer 2; Store at 4°C
• Buffer G; Store at 4°C
• Purification Columns; Store at RT
• DEPC Water; Store at RT

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