Description

Nucleic acid testing (NAT) is the method of choice for detection and quantification of a wide range of micro organisms. Primerdesign manufactures and supplies high quality quantitative real-time PCR kits for the detection and simultaneous quantification of numerous significant pathogens . A copy number standard curve is provided for quantification and an the internal extraction template (DNA or RNA), controls for the quality of the nucleic acid extraction and eliminates false negative results.

The kit is designed with the broadest possible detection profile to ensure that all clinically relevant strains and subtypes are detected. Target sequences are selected by working with data from key opinion leaders in the field. Multiple sequence alignments and unprecedented real-time PCR expertise in design and validation ensure the best possible kit. Details of the target and priming specificity are included in the individual handbooks above.

Packaged, optimised and ready to use. Expect Better Data.

Exceptional value for money
Rapid detection of all clinically relevant subtypes
Positive copy number standard curve for quantification
Highly specific detection profile
High priming efficiency
Broad dynamic detection range (>6 logs)
Sensitive to < 100 copies of target Accurate controls to confirm findings 150 reactions

Introduction

This product is suitable for rapid extraction of RNA (include miRNA) from tissue, cells, blood, s and other clinical samples. RNA can be used directly for RT-PCR, quantitative RT-PCR, test of virus DNA and so on.

Details

Specifications

Features	Specifications
Main Functions	Isolation total RNA（miRNA）from tissue, cell using two columns and DNase plus reagent
Applications	RT-PCR, cDNA synthesis, second generation sequencing
Products	RNA, miRNA
Purification method	Mini spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	Clinical tissues, cells, lymphocytes
Sample amount	Tissue: <20 mgCells: <5 x 106
Yield	2-50μg
Elution volume	≥30μl
Time per run	≤25 minutes

Principle

This product is based on silica column purification. Remove paraffin by Buffer DPS. Sample lysis withproteinase K digestion requires only 15 minutes. After lysis, samples are incubated at 80ºC for 15 minutes. Transfer to an adsorption column and RNA is adsorbed on the membrane, while protein is not adsorbed andis removed with filtration. After washing proteins and other impurities, RNA was finally eluted with low-saltbuffer.

Advantages

• Efficient DNA removal – one step RNA extraction can effectively remove genomic DNA
• High quality – one step RNA extraction reagent combined with silica gel column can obtain the highest concentration
• Fast – the whole extraction only takes 15-25 minutes
• Nontoxic – no toxic phenol chloroform extraction is required in the extraction

Kit Contents

Contents	IVD4121
Purification Times	50 Preps
HiPure DNA Mini Column Ⅱ	50
HiPure RNA Mini Columns	50
2ml Collection Tubes	150
Proteinase K	24 mg
Protease Dissolve Buffer	1.8 ml
DNase I	600 μl
DNase Buffer	6 ml
Buffer RTL	40 ml
RNA Digestion Buffer	15 ml
Buffer RWC*	20 ml
Buffer RW2*	20 ml
Nuclease Free Water	10 ml

Storage and Stability

Proteinase K should be stored at 2–8°C upon arrival. DNase I should be stored at -20°C. However, short-term storage (DNase I up to 1 weeks, Proteinase K up to 8 weeks) at room temperature (15–25°C) does not affect their performance. The remaining kit components can be stored at room temperature (15–25°C) and are stable for at least 18 months under these conditions.

This product is suitable for rapid extraction of RNA (include miRNA) from tissue, cells, blood, s and other clinical samples. RNA can be used directly for RT-PCR, quantitative RT-PCR, test of virus DNA and so on.

Overview

• Purification and enrichment of intact urinary exosomes for functional studies
• Isolate all sizes of RNA, including microRNA, irrespective of size or GC content, without bias.
• Versatile urine input volumes
• No phenol extractions, Proteinase K treatment, nor carrier RNA required
• No time-consuming ultracentrifugation, filtration nor special syringes are required
• No precipitation reagents nor overnight incubation required
• Compatible with urine from most species
• Pure exosomes are purified and are free from any other RNA-binding proteins
• Purification is based on spin column chromatography that uses Norgen’s proprietary resin
• The purified RNA is suitable for a variety of downstream applications, including Small RNA Sequencing. Find out more information on Norgen’s NGS services

Norgen’s Exosome Purification and RNA Isolation Kits constitute all-in-one systems for the purification of exosomes and the subsequent isolation of exosomal RNA from different urine sample volumes. The purification is based on spin column chromatography that employs Norgen’s proprietary resin. These kits are designed to isolate all sizes of RNA, including microRNA, irrespective of size or GC content, without bias. These kits provide a clear advantage over other available kits in that they do not require any special instrumentation, protein precipitation reagents, extension tubes, phenol/chloroform or protease treatments. Moreover, the kit allows the user to elute into a flexible elution volume ranging from 50 µL to 100 µL. The purified RNA is free from any protein-bound circulating RNA and of the highest integrity. The purified RNA can be used in a number of downstream applications including real time PCR, reverse transcription PCR, Northern blotting, RNase protection and primer extension, and expression array assays.

Urine Exosome Purification and RNA Isolation Mini Kit

For sample volumes ranging from 250 µL to 1 mL.

Urine Exosome Purification and RNA Isolation Midi Kit

For sample volumes ranging from 2 mL to 10 mL.

Urine Exosome Purification and RNA Isolation Maxi Kit

For sample volumes ranging from 11 mL to 30 mL.

Details

Supporting Data

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Kit Specifications
Minimum Urine Input	250 μL
Maximum Urine Input	1 mL
Size of Exosomes Purified	40 nm – 150 nm
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 4 of the product insert for the average yields and the common RNA quantification methods.

Storage Conditions and Product Stability
All buffers 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.

Important Note
Urine samples stored at -70°C, -20°C or at 4°C will develop some precipitation due to the aggregation of some of the highly abundant proteins in urine. Eliminating these precipitates using centrifugation or filtration may cause the loss of exosomes. Furthermore, these precipitates may affect the quality of the purified nucleic acid. We recommend the use of Norgen’s Urine Preservative when collecting urine samples, which is designed for the preservation of nucleic acids and proteins in fresh urine samples at ambient temperatures. The components of the Urine Preservative allow samples to be stored for over 2 years at room temperature with no detected degradation of urine DNA, RNA or proteins. Norgen’s Urine Preservative is available as a liquid format in Norgen’s Urine Preservative Single Dose Ampules, as well as in a dried format in Norgen’s Urine Collection and Preservation Tubes.

Component	Cat. 58400 (50 preps)	Cat. 58700 (25 preps)	Cat. 58800 (15 preps)
Slurry E	12.5 mL	12.5 mL	12.5 mL
ExoC Buffer	8 mL	30 mL	50 mL
ExoR Buffer	12 mL	12 mL	12 mL
Lysis Buffer A	20 mL	20 mL	20 mL
Lysis Additive B	2 mL	2 mL	2 mL
Wash Solution A	18 mL	18 mL	18 mL
Elution Solution A	6 mL	6 mL	6 mL
Mini Filter Spin Columns	50	25	15
Mini Spin Columns	50	25	15
Collection Tubes	50	25	15
Elution Tubes (1.7 mL)	100	50	30
Product Insert	1	1	1