Overview

• CE-IVDR marked in accordance with the European Commission Regulation (EU) No. 2017/746.
• Ideal for use in in vitro diagnostic workflows
• Simultaneous isolation of both host DNA and microbial DNA  (universal protocol)
• Eliminates PCR inhibitors including humic acids
• Fully compatible with Norgen’s Stool Nucleic Acid Collection and Transport Tubes
• High quality DNA for sensitive downstream applications including PCR, qPCR, sequencing and microarray​

This kit provides a convenient and rapid method to isolate total DNA from fresh, frozen and preserved stool samples, including those preserved using Norgen’s Stool Nucleic Acid Collection and Preservation Devices Dx (Cat. Dx45660). The universal protocol conveniently allows for the isolation of total genomic DNA from all the various microorganisms and host cells found in the stool sample simultaneously. The kit removes all traces of humic acids and other inhibitors using Bead Tubes and a combination of chemical and physical homogenization and lysis, without the need of any phenol-chloroform extractions. A simple and rapid spin column procedure is then used to further purify the DNA with high yields and molecular weights of up to 50 kb plus.  The purified DNA is of the highest quality and is fully compatible with all downstream applications such as PCR, qPCR, NGS and microarrays since all humic acid substances and other PCR inhibitors are removed during the isolation process.

NOTE: This product is not available for sale in the United States.

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	Frozen, fresh or preserved stool
Format	Spin Column
Maximum Column Binding Capacity	50 μg
Maximum Column Loading Volume	650 μL
Elution Volume	50 μL
Time to Complete 10 Purifications	30 minutes
Applications	PCR, qPCR, Southern Blot Analysis, Sequencing, Microarray Analysis.

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. Dx27600 (50 preps)
Lysis Solution	60 mL
Lysis Additive	6 mL
Binding Solution	7 mL
Wash Solution I	30 mL
Wash Solution II	22 mL
Elution Buffer	3 mL x 2
Bead Tube	50
Mini Spin Columns	50
Collection Tubes	50
Elution Tubes (1.7 mL)	50
Product Insert	1

DBCO-PEG4-NHS Ester is a click chemistry PEG reagent containing NHS ester that is able to react specifically and efficiently with primary amines (e.g. the side chain of lysine residues or aminosilane-coated surfaces) at neutral or slightly basic condition to form a covalent bond. The hydrophilic PEG spacer arm improves water solubility and provides a long and flexible connection that minimizes steric hindrance involved with ligation. DBCO is commonly used for copper-free Click Chemistry reactions. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

DBCO-PEG4-NHS Ester is a click chemistry PEG reagent containing NHS ester that is able to react specifically and efficiently with primary amines (e.g. the side chain of lysine residues or aminosilane-coated surfaces) at neutral or slightly basic condition to form a covalent bond. The hydrophilic PEG spacer arm improves water solubility and provides a long and flexible connection that minimizes steric hindrance involved with ligation. DBCO is commonly used for copper-free Click Chemistry reactions. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

Introduction

The Kit provides fast purification of high-quality RNA from paxigene Blood RNA Tube using silica-membrane spin columns. RNA purified using the HiPure System is ready for applications such as RT-PCR, Northern blotting, poly A+ RNA (mRNA) purification, nuclease protection, and in vitro translation.

Details

Specifications

Features	Specifications
Main Functions	Isolation total RNA from Paxgene RNA Tubes
Applications	RT-PCR, qPCR, Northern hybridization, NGS, nucleic acid protection, in vitro translation
Purification method	Mini spin column
Purification technology	Silica technology, DNA filtration technology
Process method	Manual (centrifugation or vacuum)
Sample type	Blood in the preservation tube
Sample amount	2.5ml
Elution volume	≥30μl
Liquid carrying volume per column	800µl
Binding yield of column	100μg

Principle

The Kit is for the purification of total RNA from 2.5 ml human whole blood collected in a PAXgene Blood RNA Tube. Purification begins with a centrifugation step to pellet nucleic acids in the PAXgene Blood RNA Tube. The pellet is washed and resuspended, and incubated in optimized buffers together with proteinase K. DNA wash The lysate is passed through a DNA Mini column. Ethanolis added to adjust binding conditions, and the lysate is applied to a column.RNA is selectively bound to the silica membrane as contaminants pass through. Remaining contaminants are removed in several efficient wash steps. Between the first and second wash steps, the membrane is treated with DNase I to remove trace amounts of bound DNA. After the wash steps, RNA is eluted in elution buffer and heat-denatured.

Advantages

• High quality – high purity total RNA can be directly used in various sensitive downstream applications
• Fast – isolation of several samples can be completed in 40 minutes by using column purification method
• Safety – no phenol chloroform extraction required
• Sensitive – direct lysis of blood, plasma and other samples without separation of leukocytes

Kit Contents

Contents	R416802	R416803
Purification Times	10 Preps	50 Preps
HiPure RNA Mini Columns I	10	50
gDNA Filter Mini Columns	10	50
2ml Collection Tubes	30	150
RNase Free Water	60 ml	250 ml
Buffer MBR1	10 ml	30 ml
Buffer MBR2	5 ml	15 ml
Buffer RW1	15 ml	60 ml
Buffer RW2*	6 ml	20 ml
Proteinase K	12 mg	50 mg
Protease Dissolve Buffer	1.8 ml	5 ml
DNase I	120 µl	600 µl
DNase Buffer	6 ml	30 ml

Storage and Stability

Proteinase K should be stored at 2–8°C upon arrival. DNase I should be stored at -20°C. However, short-term storage (DNase I up to 1 weeks, Proteinase K up to 8 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

The Kit provides fast purification of high-quality RNA from paxigene Blood RNA Tube using silica-membrane spin columns. RNA purified using the HiPure System is ready for applications such as RT-PCR, Northern blotting, poly A+ RNA (mRNA) purification, nuclease protection, and in vitro translation.