Norgen’s EXTRAClean Urine Cell-Free Circulating RNA Purification Kits provide a fast, reliable, reproducible, and simple procedure for isolating circulating RNA and exosomal RNA from various urine inputs ranging from 250 μL and up to 30 mL. The purified RNA is of the highest integrity, and can be used in a number of downstream applications including real-time PCR, reverse transcription PCR, Northern blotting, RNase protection and primer extensions, and expression array assays.
EXTRAClean Cell-Free Circulating RNA Purification Mini Kit: For sample volumes ranging from 250 μL to 2 mL
EXTRAClean Cell-Free Circulating RNA Purification Midi Kit: For sample volumes ranging from 2mL to 10mL. The first column will handle the large volume input of urine that is followed by a concentration on a mini column for a final elution of 50 μL to 100 μL
EXTRAClean Cell-Free Circulating RNA Purification Maxi Kit: For sample volumes ranging from 10 mL to 30 mL. The first column will handle the large volume input of urine that is followed by a concentration on a mini column for a final elution of 50 μL to 100 μL.
All sizes, including miRNA and small RNA (<200 nt)
Average Yields ¥
Variable depending on specimen
¥Please check page 5 of the protocol for average urine yields and common RNA quantification methods.
Storage Conditions and Product Stability
All buffers should be kept tightly sealed and stored at room temperature. These kits are stable for 2 years after the date of shipment. It is recommended to warm Lysis Buffer A for 20 minutes at 60°C if any salt precipitation is observed.
The Heat&Run® gDNA removal kit removes contaminating gDNA from RNA prior to RT-qPCR.
The kit is based on the recombinant heat labile dsDNase, which is irreversibly inactivated at low temperatures (5 minutes at 58ºC or 15 minutes at 55ºC).
Reverse Transcription can be performed in the same tube, thereby minimizing pipetting steps and reducing hands-on time.
Protocol
The kit contains HL-dsDNase which efficiently removes gDNA from RNA preps before running RT-qPCR.
gDNA is removed, leaving RNA ready for reverse transcription in the same tube (Figure 1).
Heat-labile dsDNase can easily be inactivated.
This procedure minimizes pipetting steps and reduces hands-on time.
The Heat&Run kit is especially well suited for high throughput experiments.
The high activity of the HL-dsDNase at low temperature and its heat-lability makes it ideal to use in a combination with a heat activated RT enzyme. The contaminating genomic DNA is cleaved at ambient temperature and increasing the temperature over 50°C will inactivate the DNase and start the Reverse Transcriptase.
Do you require gDNA removal in applications other than RT-qPCR? Contact our support team for assistance in implementing dsDNase treatment in your workflow.
Kit Contents
10X reaction Buffer
HL-dsDNase (50 or 250 reactions)
Document
The Heat&Run® gDNA removal kit removes contaminating gDNA from RNA prior to RT-qPCR.
The kit is based on the recombinant heat labile dsDNase, which is irreversibly inactivated at low temperatures (5 minutes at 58ºC or 15 minutes at 55ºC).
Reverse Transcription can be performed in the same tube, thereby minimizing pipetting steps and reducing hands-on time.
