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

This product uses an improved salt precipitation purification method to provide a safe and economical solution for High Weight genomic DNA extraction from blood samples, tissue samples, cultured cells, oral swabs, bacteria, and other samples. The extraction does not require the use of toxic phenol chloroform or any expensive reagents, making it the most economical reagent kit for nucleic acid extraction at present. This kit has no limit on the amount of sample used and can flexibly adjust various amounts of samples. The obtained DNA can be directly used for experiments such as PCR, enzyme digestion, Southern hybridization, and the Third-generation sequencing.

Details

Principles

SolPure DNA Kits is an improved salt precipitation purification method. (Blood samples are lysed in red blood cell lysis buffer to remove red blood cells, and white blood cells are collected by centrifugation.) After lysis, DNA is released into the lysis buffer. RNA is removed by RNASE A. High salt solution is added to precipitate proteins and impurities. Centrifuge to remove precipitate and obtain supernatant containing only DNA. Add isopropanol to precipitate and recover DNA. Wash with 70% ethanol to remove salt, and finally add Buffer TE to dissolve DNA.

Kit Contents

Contents	D3317-01	D3317-02	D3317-03
Purification Times	10	50	250
10 x Buffer RBC	4 ml	50 ml	100 ml
Buffer STE	60 ml	250 ml	2 x 550 ml
Buffer SDS (20%)	6 ml	25 ml	100 ml
Buffer PPS	20 ml	90 ml	400 ml
Proteinase K	12 mg	50 mg	240 mg
Protease Dissolve Buffer	1.8 ml	10 ml	20 ml
RNase A	5 mg	20 mg	60 mg
Buffer TE	10 ml	60 ml	250 ml

Storage and Stability

RNase A and Proteinase K should be stored at 2–8°C upon arrival. However, short-term storage (up to 4 weeks) at room temperature (15–25°C) does not affect its performance. The remaining kit components can be stored dry at room temperature (15–25°C) and are stable for at least18 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.

This product uses an improved salt precipitation purification method to provide a safe and economical solution for High Weight genomic DNA extraction from blood samples, tissue samples, cultured cells, oral swabs, bacteria, and other samples. The extraction does not require the use of toxic phenol chloroform or any expensive reagents, making it the most economical reagent kit for nucleic acid extraction at present. This kit has no limit on the amount of sample used and can flexibly adjust various amounts of samples. The obtained DNA can be directly used for experiments such as PCR, enzyme digestion, Southern hybridization, and the Third-generation sequencing.

Details