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 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 applications

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 Transport Devices Dx (Cat. Dx45660). 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.

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
Type of Stool Processed	Frozen and fresh stool
Maximum Column Binding Capacity	50 μg
Maximum Column Loading Volume	650 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. Dx49500 (50 preps)
Lysis Solution	60 mL
Wash Solution	22 mL
Elution Buffer	6 mL
Bead Tubes	50
Spin Columns	50
Collection Tubes	50
Elution Tubes (1.7 mL)	50
Product Insert	1

Overview

• Well-accepted microRNA sequence used for normalization in gene expression studies
• Best suited for RNA purification from samples with low RNA abundance including liquid biopsies (plasma/serum/urine etc.) and low cell or tissue inputs
• Compatible to expression analysis using RT-qPCR – both RNA and matching forward PCR primer provided.
• Fully compatible to Norgen’s microScript cDNA Synthesis system
• Fully compatible to Next Generation Sequencing (Small RNA-Seq) library preparation workflow

The amount of RNA that can be extracted from different biological or clinical samples varies greatly. For example, while a few micrograms of RNA could be easily purified from tissues and cells in excess amounts (such as from a few milligrams of tissue), many liquid biopsy samples may yield very low amounts of RNA. In fact, samples such as urine or plasma may yield 1 – 100 ng or less RNA per 100 µL of sample. Such a range of RNA quantity is often below the detection limit of most commonly used techniques for measuring RNA including nano-spectrophotometry and fluorescent nucleic acid stains. As a result, without properly determined RNA concentration, it becomes very difficult to normalize the starting quantity of RNA used in gene expression studies.

Norgen’s microRNA (cel-miR-39) Spike-In Kit offers a quantified synthetic RNA (cel-miR-39) for spike-in during RNA extraction procedures and subsequent normalization in RT-qPCR assays. The amount of cel-miR-39 RNA recovered after RNA extraction is directly correlated with the amount of total RNA recovered. After reverse transcription (such as with Norgen’s microScript Reverse Transcription system) of the sample RNA (with spike-in), the level of cel-miR-39 can be determined by subjecting the cDNA generated to quantitative PCR (qPCR) using fluorescent nucleic acid stains such as SYBR Green.  A cel-miR-39 specific primer is included in the kit. The level of expression of any target transcripts in different RNA samples can now be normalized to the cel-miR-39 transcript level using standard method such as ∆∆Ct relative quantification.

In addition, the cel-miR-39 RNA is compatible to library preparation methods (including ligation-based protocols) in Next Generation Sequencing (Small RNA-Seq) workflows. The cel-miR-39 RNA could be used for normalization as well as for tracking library construction efficiency.

Details

Supporting Data

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Storage Conditions
Upon receipt, store Norgen’s microRNA (cel-miR-39) Spike-In Kit at -20°C or lower. Avoid multiple freeze-thaw cycles. If needed, prepare smaller working aliquots and store at -20°C or lower.

Component	Cat. 59000
cel-miR-39 RNA	10 pmol
cel-miR-39 Forward PCR Primer	1 nmol
Nuclease-Free Water	1.25 mL
Product Insert	1

Introduction

The Kit is specially designed for simultaneous purification of genomic DNA and total RNA from formalin-fixed, paraffin-embedded (FFPE) tissue sections. The Purified DNA/RNA is used for RT-PCR and PCR detection.

Details

Specifications

Features	Specifications
Main FunctionsC	Co-isolation DNA and RNA from a single FFPE tissue sample
Applications	RT-PCR, cDNA synthesis, PCR and second-generation sequencing, etc.
Purification method	Mini spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	FFPE slice, FFPE embedded tissue
Sample amount	No more than six 10µm sections of 150mm2 surface area or three 20µm sections of 150mm2 surface area.

Principle

FFPE samples are incubated in an optimized lysis buffer, which results in the release of RNA and precipitation of DNA. After centrifugation, the RNA-containing supernatant and DNA-containing pellet are then processed separately to purify RNA and DNA. For RNA purification, transfer RNA Lysate to an adsorption column and RNA is adsorbed on the membrane, while protein is not adsorbed and is removed with filtration. After washing proteins and other impurities, RNA was finally eluted with low-salt buffer. For DNA purification, transfer DNA Lysate to an adsorption column and DNA is adsorbed on the membrane, while protein is not adsorbed and is removed with filtration. After washing proteins and other impurities, DNA was finally eluted with low-salt buffer.

Advantages

1.Use non-toxic dewaxing solution without contact with xylene
2.Obtain both DNA and RNA simultaneously from the same sample. Elute separately without affecting each other (Have the same steps and effects as top brand 80234, perfect substitute.)

Kit Contents

Contents	IVD5116
Purification Times	50 Preps
HiPure DNA Micro Column	50
HiPure RNA Mini Column I	50
2ml Collection Tubes	150
Proteinase K	50 mg
Protease Dissolve Buffer	5 ml
Buffer DPS	60 ml
Buffer FRL	15 ml
Buffer ATL	15 ml
Buffer RLC	15 ml
Buffer AL	15 ml
Buffer VHB	44 ml
Buffer RW2	25 ml
RNase Free Water	10 ml
Buffer AE	10 ml

Storage and Stability

Proteinase K 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

The Kit is specially designed for simultaneous purification of genomic DNA and total RNA from formalin-fixed, paraffin-embedded (FFPE) tissue sections. The Purified DNA/RNA is used for RT-PCR and PCR detection.