Overview

• Simultaneous isolation of both host RNA and microbial RNA (universal protocol)
• Isolate full diversity of RNA from large RNA down to small and microRNAs
• Eliminates PCR inhibitors including humic acids
• High quality RNA for sensitive downstream application
• Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix

This kit provides a convenient and rapid method to purify total RNA from small amounts of stool samples. All types of stool samples can be processed with this kit, including animal fecal samples, manure and samples collected using Norgen’s Stool Nucleic Acid Collection and Preservation Tubes (Cat. 45660). The kit removes all traces of humic acids using rapid and simple spin column procedures. Bead tubes are also provided for effective homogenization of stool. The kit purifies all sizes of RNA, from large mRNA and ribosomal RNA down to microRNA and small interfering RNA. Both host and microbial RNA is recovered. The protocol does not rely on the use of phenol or chloroform, thereby providing a user friendly procedure and allowing high-throughput analysis. The purified RNA is of the highest integrity, and can be used in a number of downstream applications including real time PCR and reverse transcription PCR for gene expression analysis. The procedure can be completed in approximately 30 minutes for 10 samples.

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Details

Supporting Data

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Kit Specifications
Maximum Stool Input	200 mg (fresh/frozen stool) or 400 μL (preserved stool)
Type of Stool Processed	Preserved, fresh, and frozen stool from humans and animals
Maximum Column Binding Capacity	50 μg
Maximum Column Loading Volume	600 μL
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. 49500 (50 preps)
Lysis Buffer C	60 mL
Wash Solution A	38 mL
Elution Buffer E	6 mL
Bead Tubes	50
Spin Columns	50
Collection Tubes	50
Elution Tubes (1.7 mL)	50
Product Insert	1

Introduction

This product is suitable for rapid extraction of DNA from FFPE sample, tissue, cells, blood, swabs, blood spots, semen and other clinical samples. This product uses size selection magnetic beads, which can selectively remove small sizes of DNA (100-300bp) from FFPE samples by adjusting the amount of binding solution, so as to improve the effective data volume of downstream NGS.

Details

Specifications

Features	Specifications
Main Functions	Isolation total DNA from FFPE using high bind beads
Applications	RT-PCR, northern blot, poly A purification, nucleic acid protection and in vitro translation, etc.
Purification technology	Magnetic beads technology
Process method	Manual or automatic
Sample type	Large quantities of solids
Sample amount	Appropriate
Elution volume	≥50μl
Time per run	30 – 120 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

1.Use non-toxic dewaxing solution without contact with xylene

2.Efficient removal of formaldehyde modification on DNA and improvement of PCR sensitivity

3. One of the best FFPE DNA extraction kits on the market, the same effect of paraffin slice extraction as top brand, and the effect of puncture sample extraction is even better than top brands

Kit Contents

Contents	D632301B	D632302B
Purification Times	48 Preps	96 Preps
MagBind Particles	1.1 ml	2 x 1.1 ml
RNase A	10 mg	20 mg
Proteinase K	24 mg	48 mg
Protease Dissolve Buffer	3 ml	6 ml
Buffer DPS	60 ml	100 ml
Buffer ATL	15 ml	30 ml
Buffer AL	15 ml	30 ml
Buffer BD*	6 ml	15 ml
Buffer BXW1*	13 ml	44 ml
Elution Buffer	15 ml	30 ml

Storage and Stability

RNase A, Proteinase K and MagBind Particles should be stored at 2-8°C upon arrival. However, short-term storage (up to 12 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.

Experiment Data

This product is suitable for rapid extraction of DNA from FFPE sample, tissue, cells, blood, swabs, blood spots, semen and other clinical samples. This product uses size selection magnetic beads, which can selectively remove small sizes of DNA (100-300bp) from FFPE samples by adjusting the amount of binding solution, so as to improve the effective data volume of downstream NGS.

Description 

The ExcelDye™ 6× DNA Loading Dye (Green) is pre-mixed buffer for tracking the DNA sample during the electrophoresis on agarose or polyacrylamide gels. It contains two dyes (Xylene cyanol FF and Orange G) for tracking the DNA migration. The Xylene cyanol FF and Orange G migrate at approximately 800 bp and 30 bp on a standard 2% TAE agarose gel respectively (4,000 bp and 50 bp on 1% TAE agarose gel respectively). The included glycerol keeps the DNA at the bottom of the well and the presence of EDTA chelates divalent metal ions to prevent the process of metal-dependent nuclease.

Composition 

• 0.03% Xylene cyanol FF 
• 0.15% Orange G 
• 10 mM Tris-HCl (pH 8.0) 
• 60% glycerol 
• 60 mM EDTA

Storage 

4°C for 12 months
-20°C for 36 months

The ExcelDye™ 6× DNA Loading Dye (Green) is pre-mixed buffer for tracking the DNA sample during the electrophoresis on agarose or polyacrylamide gels. It contains two dyes (Xylene cyanol FF and Orange G) for tracking the DNA migration. The Xylene cyanol FF and Orange G migrate at approximately 800 bp and 30 bp on a standard 2% TAE agarose gel respectively (4,000 bp and 50 bp on 1% TAE agarose gel respectively). The included glycerol keeps the DNA at the bottom of the well and the presence of EDTA chelates divalent metal ions to prevent the process of metal-dependent nuclease.