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26
RNeasy Mini Handbook 10/2019
Eliminating genomic DNA contamination
Generally, DNase digestion is not required with RNeasy Kits since RNeasy silica-membrane
technology efficiently removes most of the DNA without DNase treatment. However, further
DNA removal may be necessary for certain RNA applications that are sensitive to very small
amounts of DNA (e.g., TaqMan RT-PCR analysis with a low-abundance target). In these cases,
residual DNA can be removed by optional on-column DNase digestion using the RNAse-Free
DNase Set (see Appendix D, page 82). The DNase is efficiently removed in subsequent wash
steps. Alternatively, residual DNA can be removed by a DNase digestion after RNA
purification (see Appendix E, page 85). The DNase digestion can then be cleaned up, if
desired, using “Protocol: RNA Cleanup” (page 67).
The RNeasy Plus Mini Kit, which is designed for RNA purification from animal cells and tissues,
integrates unique gDNA Eliminator spin columns with RNeasy technology. Genomic DNA is
effectively removed in a single, rapid centrifugation step, avoiding the need for DNase
digestion. See page 91 for ordering information.
If the purified RNA will be used in real-time, two-step RT-PCR, we recommend using the
QuantiTect
®
Reverse Transcription Kit. The kit provides a fast and convenient procedure,
enabling cDNA synthesis and genomic DNA removal in only 20 min. For ordering
information, see page 91.
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