Rnase free dnase set

Manufactured by Roche

Sourced in Germany

The RNase-Free DNase Set is a laboratory product designed for the removal of DNA contamination from RNA samples. It contains a DNase enzyme that selectively degrades DNA, ensuring the purity of RNA for downstream applications.

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Lab products found in correlation

Mobio ultraclean soil dna isolation kit, by Qiagen (1 mentions) Mobio rna powersoil total rna isolation kit, by Qiagen (1 mentions) Transcriptor high fidelity cdna synthesis kit, by Roche (1 mentions) Nanodrop nd 100 spectrometer, by Thermo Fisher Scientific (1 mentions) Rnase cocktail, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

DNase (769 citations) RNase-free DNase (112 citations) Rnase-free Dnase I Set (2 citations)

2 protocols using rnase free dnase set

Soil DNA/RNA Extraction and cDNA Synthesis

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For each soil-sample replicate, the total community-DNA was separately extracted from four 1 g subsamples using the bead-beating method by following the manufacturer’s instructions for the MoBioUltraClean Soil DNA Isolation kit (MoBio Laboratories, Solana Beach, CA, USA). The extracts were pooled and further concentrated at 35 °C to a final volume of 20 μl using a Savant Speedvac® concentrator.
Total RNA was extracted from four 2 g subsamples of each replicate following the manufacturer’s instructions for the MoBio RNA PowerSoil Total RNA Isolation kit (MoBio Laboratories, Solana Beach, CA, USA). To remove residual DNA, DNase I enzyme was added using the RNase-Free DNase Set (Roche Applied Science, Penzberg, Germany) following the manufacturer’s instructions. The extracts were pooled and further concentrated at 35 °C to a final volume of 80 μl using a Savant Speedvac® concentrator. The cDNA was synthesized from 1–2 μg of total DNase-treated RNA using the Transcriptor High Fidelity cDNA Synthesis Kit according to the manufacturer’s instructions (Roche Applied Science, Penzberg, Germany). The synthesis reaction was carried out at 50 °C for 30 min. The concentration and quality of the final DNA/RNA/cDNA samples were checked with the aid of a Nanodrop® ND-100 spectrometer (Nanodrop Technologies, Wilmington, DE, USA).

Moreno B, & Benitez E. (2016). Impact of agricultural management on bacterial laccase-encoding genes with possible implications for soil carbon storage in semi-arid Mediterranean olive farming. PeerJ, 4, e2257.

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Efficient EV Purification via Centrifugation and Enzymatic Digestion

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Samples were processed by first centrifuging them at 2,000 × g for 10 minutes at room temperature in a 1.5 mL microcentrifuge tube (total recovered volume per subject was about 1.5 mL); this step was to pellet cellular debris. The supernatant was collected via pipette (cellular debris left behind) and transferred to a new 1.5 mL microcentrifuge tube, and the pellets were discarded.
The supernatant was then treated with RNase and DNase to remove any RNA or DNA that may be associated with the outside of the EVs.^{2 , 36 (link)} The single RNase and DNase treatment was performed by subjecting samples to (1 mL final volume with enzymes) 100 μL of DNA digest buffer (Qiagen, RNase-Free DNase Set; product number: 1023460), 100 μL of DNase1 (Roche, 10 mg/mL; product number: 112849320001), and 180 μL of RNase (Invitrogen RNase cocktail, 50 U/ml RNase A, 20,000 U/ml RNase T1; product number: AM2286). Samples were then incubated at 37°C for 30 minutes to allow the enzymes to act while at the same time keeping the general conditions across samples constant; the samples were then stored on ice for 15 minutes to significantly reduce further RNase/DNase enzymatic activity during the process of EV isolation.

Pucker A.D., Ngo W., Postnikoff C.K., Fortinberry H, & Nichols J.J. (2022). Tear Film miRNAs and their Association with Human Dry Eye Disease. Current eye research, 47(11), 1479-1487.

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