Overview

• Isolate high quality total RNA (including small RNA and microRNA) and all sizes of circulating and exosomal RNA, including microRNA from urine and cerebrospinal fluid (CSF) samples
• Small urine and CSF input ranging from as low as 0.5 mL to 1 mL
• No phenol extractions
• Very sensitive and linear down to a few cells without the need for carrier RNA
• Bind and elute all RNA irrespective of size or GC content, without bias
• Rapid and convenient spin column protocol
• Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix

Norgen’s Urine microRNA Purification Kit provides a rapid method for the isolation and purification of small RNA molecules (All sizes, including < 200 nt) from urine samples. These small RNAs include regulatory RNA molecules such as microRNA (miRNA) as well as tRNA and 5S rRNA. Small RNA molecules are often studied due to their ability to regulate gene expression. Typically miRNAs are 20-25 nucleotides long and regulate gene expression by binding to mRNA molecules and affecting their stability or translation. Several recent studies have shown that miRNA regulates cell growth and apoptosis. Furthermore, clinical and experimental analyses suggested that miRNAs may function as a novel class of oncogenes or tumor suppressor genes. MicroRNA expression profiles of different tumor types, relative to their normal tissues, have recently been shown to provide phenotypic signatures for particular cancer types. Unique patterns of aberrant miRNA expression may serve as molecular biomarkers for tumor diagnosis, prognosis of disease-specific outcomes, and prediction of therapeutic responses.

Details

Supporting Data

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Kit Specifications
Volume of Urine Processed	0.5 – 1 mL
Size of RNA Purified	All sizes, including < 200 nt
Time to Complete 10 Purifications	< 30 minutes

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. 29000 (25 preps)
Lysis Buffer A	2 x 20 mL
Wash Solution A	18 mL
Elution Solution A	6 mL
Mini Spin Columns	25
Collection Tubes	25
Elution Tubes (1.7 mL)	25
Product Insert	1

Introduction

A rapid, high performance dye-terminator removal process based on the paramagnetic bead technology. The paramagnetic bead format requires no centrifugation or filtration and is easily performedmanually or fully automated for high throughput dye-terminator removal. Compared to similar systems, this product produces sequences with longer Phred 20 read lengths and higher signal intensities than any other purification technology.

Details

Specifications

Features	Specifications
Main Functions	Removal of free fluorescent dye from sequencingsolution (Replace Beckmen or agencourt CleanSeq)
Applications	Automated sequences
Purification technology	Magnetic beads technology
Process method	Manual or automatic
Sample type	DNA doped with fluorescentdye
Sample amount	5μl
Recovery	90%
Elution volume	≥25μl
Operation time	≤50 minutes

Principle

The CleanSeq method contains magnetic particles in an optimized binding buffer to selectively capture sequencing extension products. The protocol can be performed directly in the thermal cycling plate. Unincorporated dyes, nucleotides, salts and contaminants are removed using a simple washing procedure. The purification procedure is amenable to a variety of automation platforms since it requires no centrifugation or vacuum filtration.

Advantages

• High recovery – up to 90%
• High throughput – using magnetic beads purification technology
• Simple – simplified operation process, combination – washing – elution

Kit Contents

Contents	BCS-5	BCS-50	BCS-500
CleanSeq Beads	5 ml	50 ml	500 ml

Storage and Stability

CleanSeq Beads should be stored at 2-8°C upon arrival and is stable up to 6 months under the condition. However, short-term storage (up to 12 weeks) at room temperature (15-25°C) does not affect its performance. Mix CleanSeq Beads well before using. The reagent should appear homogenous and consistent in color.

DO NOT FREEZE.

A rapid, high performance dye-terminator removal process based on the paramagnetic bead technology. The paramagnetic bead format requires no centrifugation or filtration and is easily performedmanually or fully automated for high throughput dye-terminator removal. Compared to similar systems, this product produces sequences with longer Phred 20 read lengths and higher signal intensities than any other purification technology.

Introduction

This product provides fast and easy methods for purification of total DNA for reliable PCR and southern blotting. Total DNA(e.g., genomic, viral, mitochondrial) can be purified from whole blood, plasma, serum, buffy coat, bone marrow, other body fluids, lymphocytes, cultured cells.

Relevant reagents were pre-packed in 96-well plates in accordance with the optimal protocol.

Details

Specifications

Features	Specifications
Main Functions	Isolation total DNA from 200μl whole blood
Applications	PCR, southern bolt and virus detection, etc
Purification technology	Magnetic beads technology
Process method	Manual or automatic
Sample type	Anticoagulant blood, concentrated blood, buffy coat, lymphocytes and cultured cells
Sample amount	200μl
Elution volume	≥50μl
Time per run	≤60 minutes

Principle

This product is based on the purification method of high binding magnetic particles. The sample is lysed and digested under the action of lysate and Protease. DNA is released into the lysate. After adding magnetic particles and binding solution, DNA will be adsorbed on the surface of magnetic particles, and impurities such as proteins will be removed without adsorption. The adsorbed particles were washed with washing solution to remove proteins and impurities, washed with ethanol to remove salts, and finally DNA was eluted by Elution Buffer.

Advantages

• High binding force – suitable for handling DNA rich samples, such as whole blood, buffy coat, concentrated blood, etc.
• Fast – polydisperse magnetic beads, fast magnetic response and short extraction time
• High purity – the obtained DNA can be directly used for second-generation sequencing, PCR based detection, gene bank, etc.
• Automatic – saving time andlabor and safer

Kit Contents

Contents	D631101	D631102	D631103
Purification Times	48	96	480
MagPure Particles	1.2 ml	2.5 ml	11 ml
Proteinase K	24 mg	50 mg	220 mg
Protease Dissolve Buffer	1.8 ml	5 ml	15 ml
Buffer AL	15 ml	30 ml	120 ml
Buffer BD	5 ml	10 ml	50 ml
Buffer GW1	22 ml	53 ml	220 ml
Elution Buffer	15 ml	30 ml	100 ml

Storage and Stability

Proteinase K, MagPure Particles 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 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 provides fast and easy methods for purification of total DNA for reliable PCR and southern blotting. Total DNA(e.g., genomic, viral, mitochondrial) can be purified from whole blood, plasma, serum, buffy coat, bone marrow, other body fluids, lymphocytes, cultured cells.

Relevant reagents were pre-packed in 96-well plates in accordance with the optimal protocol.