Introduction

The HiPure Liquid RNA & miRNA Kit integrates phenol/guanidine-based sample lysis and silica membrane purification of total RNA. MagZol LS Reagent, included in the kits, is a monophasic solution of phenol and guanidine thiocyanate designed to facilitate lysis of liquid sample and inhibit RNase. The high lysis efficiency of the reagent and the subsequent removal of contaminants by organic phase extraction enable the use of up to 0.25ml liquid sample per mini spin column.

Details

Specifications

Features	Specifications
Main Functions	Isolation total RNA from 0.25ml body fluids using column and MagZol LS reagent
Applications	qPCR / RT-PCR, liquid or solid-phasechip analysis, hybridization and SNP detection
Purification method	Mini spin column
Purification technology	Silica technology, acidphenol / guanidine extraction technology (MagZol pretreatment technology)
Process method	Manual (centrifugation or vacuum)
Sample type	Blood, serum, plasma
Sample amount	250μl
Elution volume	≥20μl
Liquid carrying volume per column	800µl
Binding yield of column	100μg

Principle

0.25ml liquid samples are homogenized in 0.75ml MagZol LS Reagent. After addition of chloroform, the homogenate is separated into aqueous and organic phases by centrifugation. The upper, aqueous phase is extracted, and ethanol is added to provide appropriate binding conditions. The sample is then applied to the spin column, where the total RNA (up to 100 µg) binds to the membrane and phenol and other contaminants are efficiently washed away. High-quality RNA is then eluted in 30–100µl of RNase-free water.

Advantages

• High quality – high purity total RNA can be directly used in various sensitive downstream applications
• Fast – isolation of several samples can be completed in 40 minutes by using column purification method
• Safety – no phenol chloroform extraction required
• Sensitive – direct lysis of blood, plasma and other samples without separation of leukocytes

Kit Contents

Contents	R416302	R416303
Purification Times	50 Preps	250 Preps
HiPure RNA Mini Columns	50	250
2ml Collection Tubes	50	2 x 125
MagZol LS Reagent	60 ml	270 ml
Buffer RWC	20 ml	60 ml
Buffer RW2*	20 ml	2 x 50 ml
RNase Free Water	10 ml	30 ml

Storage and Stability

MagZol LS Reagent should be stored at 2-8°C upon arrival. However, short-term storage (up to 24 weeks) at room temperature (15-25°C) does not affect their performance. The remaining kit components can be stored dry at room temperature (15-25°C) and are stable for at least 18 months under these conditions.

The HiPure Liquid RNA & miRNA Kit integrates phenol/guanidine-based sample lysis and silica membrane purification of total RNA. MagZol LS Reagent, included in the kits, is a monophasic solution of phenol and guanidine thiocyanate designed to facilitate lysis of liquid sample and inhibit RNase. The high lysis efficiency of the reagent and the subsequent removal of contaminants by organic phase extraction enable the use of up to 0.25ml liquid sample per mini spin column.

Overview

• Isolate all sizes of circulating and exosomal RNA, including microRNA
• Versatile plasma/serum input ranges
• 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
• High quality, purified RNA is suitable for a variety of downstream applications, including Small RNA Sequencing. Find out more information on Norgen’s NGS services
• Compatible with Streck Cell-Free RNA BCT® Tubes
• Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix

These kits provide 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 convenient spin column method. These kits can purify RNA from fresh or frozen serum or plasma samples 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. 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.

Plasma/Serum RNA Purification Mini Kit

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 50 µL to 200 µL. The purified plasma/serum RNA is eluted in a flexible final volume of 10 µL to 25 µL.

Plasma/Serum RNA Purification Midi Kit

This utilizes a two column method, and 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. 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.

Plasma/Serum RNA Purification Maxi Kit

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 2 mL to 5 mL. 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.

Isolate RNA after Purifying EVs and Exosomes

For Ultracentrifugation, Exoquick, and Filtration

Cat. #	Name	Elution Volume	Plasma/Serum	Urine	Cell-Culture Media
55000	Plasma/Serum RNA Purification Mini Kit	10 – 25 µL	50 µL – 1 mL	250 µL – 1 mL	5 – 10 mL
35300	Total RNA Purification Micro Kit	20 – 50 µL	1 – 4 mL	2 – 10 mL	10 – 20 mL
17200	Total RNA Purification Kit	50 – 100 µL	4 – 10 mL	11 – 30 mL	20 – 35 mL

Details

Supporting Data

Figure 1 / 13

Previous

Next

Click for expanded view

Kit Specifications
Sample Volume Range	50 to 200 μL
Anti-coagulant (for Plasma)*	EDTA or Citrate
Size of RNA Purified	All sizes, including small RNA (< 200 nt)
Minimum Elution Volume	10 μL
Maximum Elution Volume	25 μL
Time to Complete 10 Purifications	15-20 minutes
Average Yield**	Variable depending on specimen

*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.

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

Storage Conditions and Product Stability

All solutions should be kept tightly sealed and stored at room temperature. This kit is 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.

Kit Components	Cat. 55000 (50 preps)	Cat. 56100 (20 preps)	Cat. 56200 (10 preps)
Lysis Buffer A	2 x 20 mL	100 mL	1 x 130 mL 1 x 30 mL
Wash Solution A	18 mL	1 x 38 mL 1 x 18 mL	38 mL
Elution Solution A	6 mL	6 mL	6 mL
Elution Buffer F	–	15 mL	15 mL
Micro Spin Columns	50	–	–
Mini Spin Columns	–	20	10
Midi Spin Columns	–	20	–
Maxi Spin Columns	–	–	10
Collection Tubes	50	20	10
Elution Tubes (1.7 mL)	50	20	10
Product Insert	1	1	1

Description

Our SNPsig® kits use our own proprietary genotyping method to enable the identification of SARS-CoV-2 variants of concern. These products can be used on any real-time PCR machine using familiar protocols, whilst resulting in exceptional genotyping data.

Positive control templates for wild-type and variants are supplied in every kit to make data interpretation simple.

Our SNPsig® technology provides an alternative to sequencing as well as S gene target failure (SGTF) that enables scientists to analyse and monitor these specific genomic mutations.  Our kits can provide a pivotal role in screening for SARS-CoV-2 variants for the purpose of genomic surveillance and studies.

For the detection of the SARS-CoV-2 (20H/501Y.V2 South Africa)
Rapid detection of specific detection profiles
High priming efficiency
Sensitive to < 100 copies of target Positive copy number standard curve for quantification Accurate controls to confirm findings 96 reactions, includes master mix