Plasma/Serum Circulating and Exosomal RNA and DNA Purification Kit (Spin Columns) Mini, 10 - 200 ul sample input

Product Description

• Isolate all sizes of circulating and exosomal RNA, including microRNA
• Isolate all sizes of circulating DNA from plasma and serum samples
• Isolate viral and bacterial DNA and RNA
• Versatile plasma and serum input volumes (10 µL - 200 µL)
• No phenol extractions
• Concentrate circulating RNA, exosomal RNA and cell-free circulating DNA into a flexible elution volume ranging from 10 µL to 25 µL
• Isolate inhibitor-free nucleic acid
• Can be used to isolate genomic, viral, and bacterial DNA from milk
• Purify high-quality RNA and DNA in 30 minutes

Norgen’s Plasma/Serum RNA/DNA Purification Mini Kit provides a fast, reliable, reproducible and simple procedure for the sequential isolation of circulating RNA, exosomal RNA and Cell-Free Circulating DNA (cfc) from the same Plasma/Serum Sample. It can sequentially islated RNA and DNA from small plasma/serum input ranging from 10 µL to 200 µL. Purification is based on Norgen's new spin column chromatography that uses Norgen’s proprietary resin separation matrix. The kit is designed to isolate all sizes of circulating RNA, including microRNA, all sizes of exosomal RNA as well as all sizes of cfc-DNA from fresh or frozen plasma or serum samples. Norgen’s Plasma/Serum RNA Purification Kit provides a clear advantage over other available kits in that it does not require Phenol/Chloroform or any Protease treatments for the isolation of plasma/serum RNA or DNA . RNA and DNA can be eluted into a flexible elution volume ranging from 10 µL to 25 µL. Typical yields of free-circulating, exosomal RNA and cfc-DNA vary depending on the input sample, as the amount of RNA and/or DNA present in plasma and serum will depend upon the health status of the individual. Normally, the RNA/DNA yield from plasma or serum is highly variable (range from 1 to 100ng/mL). Variability is also observed between samples collected from the same donor at different times during the day. This kit is suitable for the isolation of RNA/DNA from serum or plasma prepared from blood collected on either EDTA or Citrate. Plasma samples prepared from blood collected on heparin should not be used as heparin can significantly interfere with many downstream applications such as RT-PCR.

Features and Benefits

• No phenol:chloroform extractions – Circulating RNA and exosomal RNA is isolated without the use of harmful chemicals such as phenol or chloroform
• Isolate all sizes of circulating RNA and exosomal RNA - The kit allows for the isolation of all sizes of fragmented circulating RNA and exosmal RNA, including microRNA
• Isolate all sizes of circulating DNA - The kit allows for the isolation of all sizes of fragmented cell-free circulating DNA, ranging from high molecular weight fragments down to fragments as small as 50bps.
• Fast and easy processing - Rapid spin column format allows for the processing of multiple samples in under 30 minutes..
• Small input volume - Isolate circulating RNA, exosomal RNA and cfc-DNA from 10 µL to 200 µL of plasma/serum.
• Concentrate circulating RNA, exosomal RNA and cfc-DNA – Circulating and exosomal RNA as well as cfc-DNA present in input volumes of 10 µL to 200 µL are concentrated into final elution volume of 10 µL to 25 µL.
• Isolate inhibitor-free nucleic acid - Purified RNA can be used in a number of sensitive downstream applications including reverse transcription qPCR, reverse transcription PCR, Northern blotting, RNase protection and primer extension, and expression array assays. Purified DNA can be used in a number of sensitive downstream applications including PCR, qPCR, methylation-sensitive PCR and Southern Blot analysis.

Applications

The purified circulating RNA and cfc-DNA is compatible with most of the molecular biological applications such as:

• Reverse transcription qPCR
• PCR
• qPCR
• Reverse transcription PCR
• Northern blotting
• Southern Blot
• RNase protection and primer extension
• Expression array assays.
• miRNA array
• RNA-Seq
• Nanostring
• Fluidigm
• Droplet/Digital PCR
• Methylation-sensitive PCR
• CpG array
• Restriction enzyme digestions
• Viral Detection
• Bacterial Detection

Data

Effective and Consistent Detection of miRNA from Plasma.

Norgen's Plasma/Serum RNA Purification Kit can effectively isolate miRNA from plasma. Circulating miRNA was isolated from 200 µL plasma using Norgen's Plasma/Serum RNA Purification Kit, miRNeasy Serum/Plasma Kit (Qiagen)and miRCURY RNA Isolation Kit – BioFluids (Exiqon). Circulating miRNA was isolated from 600 µL using mirVana™ PARIS™ Kit. Stem loop RT-qPCR using primers specific to miR-21 was performed. In brief, 1 microliter of the 15 µL purified RNA using Norgen's Plasma/Serum RNA Purification Kit, miRNeasy Serum/Plasma Kit (Qiagen) and 3.3 microliters of the 50 µL purified RNA using miRCURY RNA Isolation Kit – BioFluids (Exiqon) and mirVana™ PARIS™ Kit was then subjected to a 20 µL reverse transcription using miR-21 stem-loop reverse primer. Three microliters of the reverse transcription was used in a 20 µL real-time PCR reaction with primers to detect the human miR-21. Norgen's Plasma/Serum RNA Purification Kit is the only product that showed the most consistent and the highest recovery of the miR-21 transcripts as compared to the other isolation methods. The recovery of the miRNA from 200µL plasma was higher than that recovered from RNA purified from 600µL using mirVana™ PARIS™ Kit.

Effective and Consistent Detection of Cell-Free Circulating DNA from Plasma.

Norgen's Plasma/Serum Cell-Free Circulating DNA Purification Kit can effectively isolate Cell-Free Circulating DNA from plasma. Cell-Free Circulating DNA was isolated from 200 µL plasma using Norgen's Plasma/Serum Cell-Free Circulating DNA Purification Kit, QIAamp DNA Blood Mini Kit (Qiagen) and NucleoSpin® Blood (MACHEREY-NAGEL). Three microliters of the purified DNA was used in a 20 µL real-time PCR reaction with primers to detect the housekeeping 5S rRNA gene. Norgen's Plasma/Serum Cell-Free Circulating DNA Purification Kit is the only product that showed the most consistent and the highest recovery of the housekeeping 5S rRNA gene as compared to the other isolation methods.

Kit Specifications

Minimum Plasma/Serum Input 10 μL

Maximum Plasma/Serum Input 200 μL

Size of RNA Purified: All sizes including miRNA and small RNA (<200 nt)

Size of DNA Purified: ≥ 50 bp

Elution Volume: 10-25 μL

Time to Complete 10 Purifications: 15-20 minutes

Average Yields: Variable depending on specimen

Note: Do not exceed the recommended sample input volume of 200 µL.

Storage Conditions and Product Stability

All buffers should be kept tightly sealed and can be stored at room temperature (15-25oC) for up to 2 years without showing any reduction in performance. It is recommended to warm Lysis Buffer A for 20 minutes at 60oC if any salt precipitation is observed.