Introduction

HiPure FFPE RNA Kit supplies a simple and rapid RNA extraction for Formalin-fixed, paraffin-embedded (FFPE) tissue and sections samples. This kit is based on silica gel column purification technology, no phenol-chloroform extraction or alcohol precipitation. The whole extraction only takes 30 minutes (not including digestion time). RNA can be directly used for downstream applications such as RT-PCR, Northern blot, vitro translation and other experiments.

Details

Specifications

Principle

This product is based on silica column purification. Remove paraffin by Buffer DPS. Sample lysis with proteinase K digestion requires only 15 minutes. After lysis, samples are incubated at 80ºC for 15 minutes. Transfer 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.

Advantages

• High quality – high purity total RNA can be directly used in various sensitive downstream applications
• Fast – several samples can be extracted in 60 minutes by column method
• Safe – no phenol chloroform extraction required
• Sensitive – RNA can be recovered at the level of PG
• Efficient DNA removal – unique method to effectively remove genomic DNA

Kit Contents

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

HiPure FFPE RNA Kit supplies a simple and rapid RNA extraction for Formalin-fixed, paraffin-embedded (FFPE) tissue and sections samples. This kit is based on silica gel column purification technology, no phenol-chloroform extraction or alcohol precipitation. The whole extraction only takes 30 minutes (not including digestion time). RNA can be directly used for downstream applications such as RT-PCR, Northern blot, vitro translation and other experiments.

Overview

• Ensure optimal results for sensitive applications like NGS
• Up to a tenfold increase in microRNA mapping during sequencing runs to reduce costs
• Isolates all sizes of exosomal and extracellular vesicle RNA, including microRNA.
• Bind and elute all RNA irrespective of size or GC content, without bias.
• No phenol extractions.
• No Proteinase K treatment.
• No carrier RNA.
• Concentrate isolated RNA into a flexible elution volume ranging from 50 μL to 100 μL.
• Purify high-quality RNA in 15-20 minutes.
• Purified RNA is suitable for a variety of downstream applications, including Small RNA Sequencing.
• Purification is based on EXTRAClean spin column chromatography that uses Norgen’s proprietary separation matrix.

The kit is designed to isolate all sizes of RNA, including microRNA. The kit provides a clear advantage over other available kits in that it does not require any special instrumentation, protein precipitation reagents, extension tubes, phenol/chloroform or protease treatments. Moreover, the kit allows the user to elute into a flexible elution volume ranging from 50 μL to 100 μL. The purified RNA is of the highest integrity, and can be used in a number of downstream applications including real time PCR, reverse transcription PCR, Northern blotting, RNase protection and primer extension, and expression array assays.

Details

Supporting Data

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Storage Conditions and Product Stability
All buffers should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment. It is recommended to warm Lysis Buffer A for 20 minutes at 60°C if any salt precipitation is observed.

OverView

Applications

• Viral Bioprocessing
• Complete Removal of DNA

M-SAN HQ  – Peak performance where it matters most at physiological conditions

Medium-Salt Active Nuclease High Quality (M-SAN HQ) is a Bioprocessing Grade nuclease developed for removal of both single and double-stranded DNA and RNA at the physiological salt conditions most often used in bioprocessing and biomanufacturing workflows. M-SAN HQ allows you to directly replace Benzonase without changing your workflow.

This novel, nonspecific endonuclease is active over a broad pH range and displays optimum activity at salt concentrations between 125 – 250 mM. Due to its excellent performance at physiological conditions, M-SAN HQ can be used directly in the cell medium or the harvested supernatant without buffer adjustments. This makes M-SAN HQ ideal for the manufacturing of fragile vectors such as lentiviruses and retroviruses.

M-SAN HQ can be directly used in medium without buffer adjustments The high activity of M-SAN HQ at standard cell medium conditions leads to improved DNA clearance compared to other commonly used nucleases. In the data (see Figure 6), an over 2-fold reduction in residual DNA was achieved. 

Key Benefits

• Compatibility: Ideal for working with both fragile and robust viral vectors and proteins in a variety of cell media
• Optimum activity: Optimization for cell media salinity allows both shorter DNA fragments and reduced incubation times.
• Cost-Effectiveness: Reduced need for additional reagents and steps can lower production costs
• Quality: Maintains the integrity of sensitive or labile biological molecules, ensuring a higher quality end product
• Flexibility: Can be used directly in a variety of media without the need for customization

Key Features 

• High purity (≥ 99%)
• No protease detected
• Endotoxin-tested
• Animal origin-free production
• Supplied with extended product documentation

Adapted to use in medium without salinity adjustments 

Optimal activity at physiological salinity and pH makes it ideal for DNA removal from mammalian cell media. (see Figures)

The high activity of M-SAN HQ at the physiological salinity and pH found in standard cell medium conditions leads to improved DNA clearance compared to commonly used nucleases.

Simple to Optimize in Cell Media

M-SAN HQ is uniquely formulated to excel in physiological pH conditions, offering high performance at the commonly used cell media pH of 7.4.

While other nucleases often require more alkaline environments, M-SAN HQ stands out for its adaptability and effectiveness in cell media. It’s the nuclease that truly aligns with your bioprocessing needs. (See Figure section)

M-SAN HQ ELISA Kit

The M-SAN HQ ELISA Kit confirms the removal of M-SAN High Quality in bioprocessing and biomanufacturing applications with high accuracy: 

• 100% (±10%) Measuring range 0.47 – 7.50 ng/ml
• 100% (±20%) Measuring range 0.12 – 0.47 ng/ml

Features:

Shelf life: 18 months

Fast: 1.5 – 2 hours

Sensitive – Quantification range: 0.12 – 7.5 ng/ml

Accurate

Flexible – 96-well plate in 8-well strip format

Medium-Salt Active Nuclease High Quality (M-SAN HQ) is a Bioprocessing Grade nuclease developed for removal of both single and double-stranded DNA and RNA at the physiological salt conditions most often used in bioprocessing and biomanufacturing workflows. M-SAN HQ allows you to directly replace Benzonase without changing your workflow.