• Isolate all sizes of circulating and exosomal RNA, including microRNA
  • Versatile plasma/serum input ranging from 250 µL to 1.5 mL
  • No phenol extractions
  • No carrier RNA
  • Bind and elute all RNA irrespective of size or GC content, without bias
  • Concentrate circulating RNA and exosomal RNA into a flexible elution volume ranging from 50 µL to 100 µL
  • Purify high-quality RNA in 35-40 minutes

ItemsCat. #Size
Plasma/Serum RNA Purification Midi Kit 56100 20 preps

Plasma/Serum RNA Purification Midi Kit

This kit provides a fast, reliable and convenient method to purify and concentrate high quality, high purity and inhibitor-free cell-free circulating and exosomal RNA using a two column method. This kit can purify RNA from fresh or frozen serum or plasma samples prepared from blood collected on either EDTA or Citrate, from volumes ranging from 250 µL to 1.5 mL. 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

The first column will handle the large volume input of bodily fluids that is followed by a concentration on a mini column for a final elution of 50 µL to 100 µL. All components for the purification and concentration are provided in one convenient and fast kit for the easy processing of large input volumes of bodily fluids. For a schematic workflow of the protocol click here.

The purified plasma/serum RNA is fully compatible with all downstream applications including PCR, qPCR, methylation-sensitive reverse transcription qPCR, reverse transcription PCR, Northern blotting, RNase protection and primer extension, expression array assays, and NGS.

Background

Plasma/Serum cell-free circulating RNA or exosomal RNA has the potential to provide biomarkers for certain cancers and disease states. Exosomes are 40 - 150 nm membrane vesicles, which are secreted by most cell types. Exosomes can be found in saliva, blood, urine, amniotic fluid and malignant ascitic fluids, among other biological fluids. Evidence has been accumulating recently that these vesicles act as cellular messengers, conveying information to distant cells and tissues within the body. These exosomes may play a functional role in mediating adaptive immune responses to infectious agents and tumours, tissue repair, neural communication and transfer of pathogenic proteins. For this reason exosomal RNAs may serve as biomarkers for various diseases including cancer. As the RNA molecules encapsulated within exosomes are protected from degradation by RNAses they can be efficiently recovered from biological fluids, such as plasma or serum.

Protocol

Kit Specifications
Minimum Plasma/Serum Input
250 μL
Maximum Plasma/Serum Input
1.5 mL
Size of RNA Purified
All sizes, including miRNA
and small RNA (<200 nt)
Elution Volume
50-100 μL
Time to Complete 10 Purifications
35-40 minutes
Average Yields*
Variable depending on specimen

*Please check page 7 of the Product Insert for Average Plasma/Serum Yields and Common RNA Quantification Methods.

This kit is suitable for the isolation of RNA from fresh or frozen 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.

Storage Conditions and Product Stability
All buffers should be kept tightly sealed and 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.


Kit Components
Component Cat. 56100 (20 preps)
Lysis Buffer A 100 mL
Wash Solution A 1 x 38 mL*, 1 x 18 mL*
Elution Solution A 6 mL
Elution Buffer F 15 mL
Mini Spin Columns 20
Midi Spin Columns 20
Collection Tubes 20
Elution Tubes (1.7 mL) 20
Product Insert 1

* For the preparation of working solutions, please see the Important Notes (Notes prior to use) section in the Product Insert.

Product Information Sheets: 

Title Circulating MicroRNAs: Association with Lung Function in Asthma
Journal PLOS ONE. 2016.
Authors Kho AT, Sharma S, Davis J, Spina J, Howard D, McEnroy K, Moore K, Sylvia J, Qiu W, Weiss ST, Tantisira KG