Overview

• Purify all sizes of RNA (including microRNA) without the need for phenol
• Isolate and detect total RNA from 1 mL and up to 50 mL urine
• Provides high-quality RNA for sensitive applications – isolate RNA from as little as 100 cells
• Rapid processing time from sampling to downstream testing
• Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix

This kit provides a rapid spin column method for the isolation and purification of total RNA (including microRNA) from exfoliated cells that have been shed into the urine from the urinary tract. The kit purifies all sizes of RNA, from large mRNA and ribosomal RNA down to microRNA (miRNA) and small interfering RNA (siRNA). The RNA is preferentially purified from other cellular components such as proteins, as well as from the contaminating species found in urine such as glucose and salts, without the use of phenol or chloroform. The purified RNA is of the highest integrity and can be used in a number of downstream applications including real-time RT-PCR, qRT-PCR, Northern blotting, NGS, and expression array assays. Multiple samples can be processed in 20 minutes.

Background

RNA biomarkers from exfoliated cells can be used as non-invasive tools for a number of diagnostic and research applications including the diagnosis and monitoring of bladder, kidney, or other urinary-tract cancers. 

Details

Supporting Data

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Kit Specifications
Maximum Column Binding Capacity	50 μg
Volume of Urine Processed	1-50 mL
Maximum Input of Exfoliated Cells	1 x 106
Size of RNA Purified	All sizes, including small RNA (< 200 nt)
Time to Complete 10 Purifications	20 minutes (for Exfoliated Cells)30 minutes (for Bacteria)
Average Yield	~1μg RNA per 1 x 105 cells (For Exfoliated Cells – Varies due to cell density sample)~0.5μg RNA per 1 x 107 cells(For Bacteria – Varies due to cell density sample)

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.

Component	Cat. 22550 (25 preps)
Buffer SK	15 mL
Wash Solution A	18 mL
Elution Buffer E	6 mL
Micro Spin Columns	25
Collection Tubes	25
Elution Tubes (1.7 mL)	25
Product Insert	1

Introduction

HiPure RNA Clean Up Kit provides a rapid and easy method for the purification and concentrate RNA from enzymatic reactions or for desalting the RNA samples. RNA purified using HiPure RNA Clean Up Kit is ready for all downstream applications such as RT-PCR, Northern blotting, mRNA purification, nuclease protection, and in vitro translation.

Details

Specifications

Features	Specifications
Main Functions	Purification and concentration of RNA（mRNA>200nt or miRNA）from transcription products, DNase cleavage products, labeled products, and crude RNA products
Applications	Sequencing, ligation, enzyme digestion, RT-PCR, labeling, etc.
Purification method	Mini spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	Crude RNA, enzymatic reaction solution
Sample amount	≤100μl
Recovery	90%
Elution volume	≥15μl
Time per run	≤15 minutes
Liquid carrying volume per column	800μl
Binding yield of column	100μg

Principle

The HiPure system uses a simple bind-wash-elute procedure. Binding buffer is added directly to the sample or other enzymatic reaction, and the mixture is applied to the column. Nucleic acids adsorb to the silica membrane in the high-salt conditions provided by the buffer. Impurities are washed away and pure RNA is eluted with a small volume of low-salt buffer provided or water, ready to use in all subsequent applications.

Advantages

• High recovery – recovery of RNA up to 90%
• Low elution volume – the elution volume can be as low as 15µl
• Fast – only 10 minutes for recovery by using column method
• General – suitable for various crude RNA products
• Wide range of fragment recovery:(>25nt RNA)

Kit Contents

Contents	R214402	R214403
Purification Times	50 Preps	250 Preps
Buffer GXP	30 ml	120 ml
Buffer RW2	20 ml	2 x 50 ml
RNase-Free Water	20 ml	60 ml
HiPure RNA Mini Columns I	50	250
2 ml Collection Tubes	50	250

Storage and Stability

The Kit can be stored dry at room temperature (15-25°C) and are stable for at least 18 months under these conditions. The entire kit can be stored at 2-8°C, but in this case buffers should be redissolved before use. Make sure that all buffers are at room temperature when used.

HiPure RNA Clean Up Kit provides a rapid and easy method for the purification and concentrate RNA from enzymatic reactions or for desalting the RNA samples. RNA purified using HiPure RNA Clean Up Kit is ready for all downstream applications such as RT-PCR, Northern blotting, mRNA purification, nuclease protection, and in vitro translation.