Introduction

This kit is used for extracting total viral nucleic acid from non-cell/low cell content biological samples such as body fluid, serum, plasma, immersion solution, tissue homogenate supernatant, culture supernatant, etc., the extracted products can be used for clinical in vitro detection.

Details

Specifications

Features	Specifications
Main Functions	Extract viral RNA/DNA from 200μl plasma/serum samples
Applications	RT-PCR，PCR，NGS
Products	Viral DNA / RNA, body cell DNA / RNA
Purification method	Mini spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	Plasma, serum, effusion, urine, fecal suspension supernatant
Sample amount	200μl
Elution volume	≥30μl
Time per run
Liquid carrying volume per column	800μl
Binding yield of column	100μg

Principle

This product is based on silica gel purification. The sample is lysed and digested with lysate and protease, DNA/RNA is released into the lysate. Transfer to an adsorption plate and filter column. DNA/RNA is adsorbed on the membrane, while protein is not adsorbed and is removed with filtration. After washing proteins and other impurities, DNA / RNA was finally eluted with low-salt buffer (10 MmTris, pH 8.0).

Advantages

• Fast – column purification without PK digestion
• High quality – high purity total RNA can be directly used in various sensitive downstream applications
• Safe – no phenol chloroform extraction required
• Sensitive – DNA/RNA can be recovered at the level of PG

Kit Contents

Contents	IVD4175
Purification Times	100 Preps
HiPure Viral Column	100
2ml Collection Tubes	100
PK/Carrier RNA	50 mg/310μg
Protease Dissolve Buffer	5 ml
Buffer VLE	42 ml
Buffer CE	60 ml
RNase Free Water	15 ml

Storage and Stability

This kit is shipped and stored at room temperature and is valid for 12 months.

This kit is used for extracting total viral nucleic acid from non-cell/low cell content biological samples such as body fluid, serum, plasma, immersion solution, tissue homogenate supernatant, culture supernatant, etc., the extracted products can be used for clinical in vitro detection.

Overview

• Eight discrete bands, ranging from 100 to 1,000 bases
• Higher intensity band at 500 bases for easy reference
• No need for staining or destaining as loading dye contains ethidium bromide
• Convenient lyophilized format provides better product stability

The Norgen 100 b RNA Ladder is a set of RNA transcripts derived from recombinant DNA templates. This ladder is suitable for precise sizing of small RNA molecules using native or denaturing agarose gels, and can be easily visualized under UV.

Contents

• 2 vials of lyophilized RNA ladder (25 loads each)
• 1x loading buffer: 33.3% formamide, 11.7% formaldehyde, 0.012% bromophenol blue, 0.05mM EDTA, 0.012% ethidium bromide, and 0.7x MOPS solution
• 2x loading buffer

Details

Supporting Data

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Specifications:

• Eight discrete bands, ranging from 100 to 1,000 bases
• Higher intensity band at 500 bases for easy reference

Contents:

• 2 vials of lyophilized RNA ladder (25 loads each)
• 1x loading buffer: 33.3% formamide, 11.7% formaldehyde, 0.012% bromophenol blue, 0.05mM EDTA, 0.012% ethidium bromide, and 0.7x MOPS solution
• 2x loading buffer

Instructions:

To reconstitute the lyophilized RNA ladder, add 250 µL of 1x loading buffer to each 25 loads vial and vortex gently.
Heat at 80°C for 10 minutes. Incubate on ice for 1 min. Load 10 µL on a 1.5-2% gel. For optimal results, use Norgen 2x loading buffer with each RNA sample. There is no need for staining and destaining denaturing gels since Norgen’s loading buffer contains ethidium bromide.

Storage:

Store at -20°C.

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	11 mL
Maximum Urine Input	30 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