RNase Inhibitor

Introduction

Magen’s HiPure columns are prepared by high quality glass fiber filter membrane as raw materials through membrane cutting, membrane release, ring release, ring pressing, gland, weighing and other processes. HiPure nucleic acid adsorption columns have the characteristics of long-term stability and high binding capacity. Experiments show that the highest binding capacity and binding efficiency of HiPure nucleic acid adsorption columns are basically unchanged when stored at room temperature for 4 years.

The series of nucleic acid columns produced by Magen Biotech are based on carefully selected imported glass fiber membranes (GF/B, GF/D, GF/F). Columns production processes such as polypropylene injection molding materials, injection molding process, and downstream membrane packing and compression rings are strictly controlled. This is to ensure that the column has extremely high adsorption capacity and long-term stability. Compared with conventional products on the market, Magen’s columns are with varieties, and binding rate will not change when stored at room temperature for 4 years.

Details

Specifications

Features	Specifications
Recommended application	gDNA and RNA Isolation
Preservation conditions	Room temperature
Stability	Up to 4 years
Filter membrane	High quality glass fiber filter GF/B, 2 layers
Membrane aperture	1.0μm
Maximum binding yield of plasmid	30 μg
Maximum yield of alcohol mediated Binding	100 μg
Single liquid carrying capacity of column	900 μl
Minimum elution volume	80 μl
Withstand centrifugal force	4,000-5,000 x g
Centrifuge	Low speed centrifuge, Swing out Rotor, can placed a height of 6.5cm square,  (height of HiPure DNA Plate & 1.6ml Collection Plate: height, 6.2cm)

Adsorption Mechanism

Based on the negatively charged DNA skeleton, it has a high affinity for positively charged glass fibers. In high salt and ethanol solutions, DNA/RNA binds to glass fiber and interacts with hydrophilic matrix on silica through hydrogen bond. DNA/RNA is tightly bound. All pollutants can be removed by washing solution. At high salt concentration, nucleic acids selectively bind to silica gel membrane, while other pollutants, mainly proteins, are removed by membrane washing.

Ordering information

CAT.No.	Product Name	Package
C13131	HiPure gDNA Plate (2 x GF/B)with 1.6ml Collection Plate	10/Bag

Purchase Guide

Item No.	Product Name	Membrane type/number of layers	Collection tubes	Plasmid DNA binding capacity (Physical adsorption)	gDNA/RNA binding capacity (Alcohol-mediated adsorption)	Minimum Elution volume	Liquid volume capacity
C13010	HiPure DNA Nano Column	2 layers GF/F	2ml without cap	5μg	20μg	10μl	700μl
C13011	HiPure DNA Micro Column	3 layers GF/F	2ml without cap	10μg	50μg	15μl	700μl
C13100	HiPure DNA Mini Column I	2 layers GF/B	2ml without cap	15μg	100μg	30μl	700μl
C13110	HiPure DNA Mini Column II	4 layers GF/B	2ml without cap	35μg	200μg	50μl	800μl
C13111	HiPure RNA Mini Column	3 layers GF/B	2ml without cap	30μg	200μg	30μl	800μl
C13112	HiPure Viral Mini Column	3 layers GF/F	2ml without cap	30μg	200μg	30μl	800μl
C13113	HiPure CFDNA Mini Column	3 layers GF/F,1 layer GF/B	2ml without cap	30μg	200μg	30μl	800μl
C13120	HiPure DNA Midi Column	4 layers GF/B	15ml Centrifuge tube	125μg	1mg	500μl	4ml
C13121	HiPure DNA Midi Column III	8 layers GF/B	15ml Centrifuge tube	250μg	1mg	500μl	4ml
C13122	HiPure DNA Maxi Column	4 layers GF/B	50ml Centrifuge tube	500μg	5mg	1000μl	20ml
C13123	HiPure DNA Maxi Column III	8 layers GF/B	50ml Centrifuge tube	1mg	5mg	1000μl	20ml
C13124	HiPure DNA Maxi Column C	8 layers GF/B	50ml high speed Centrifuge tube	1mg	5mg	700μl	12ml
C13130	HiPure DNA Plate	2 layers GF/F	1.6ml Plate	30μg	100μg	80μl	900μl
C13131	HiPure gDNA Plate	2 layers GF/B	1.6ml Plate	30μg	100μg	80μl	900μl

Note: GF/B pore size is for 1.0μM glass fiber membrane; GF/F pore size is for 0.7μm glass fiber membrane.

Magen’s HiPure columns are prepared by high quality glass fiber filter membrane as raw materials through membrane cutting, membrane release, ring release, ring pressing, gland, weighing and other processes. HiPure nucleic acid adsorption columns have the characteristics of long-term stability and high binding capacity. Experiments show that the highest binding capacity and binding efficiency of HiPure nucleic acid adsorption columns are basically unchanged when stored at room temperature for 4 years.

Overview

• Isolate all sizes of RNA, including microRNA, irrespective of size or GC content, without bias
• Versatile sample input ranges
• Isolate all sizes of free-circulating RNA, including microRNA​
• Bind and elute all RNA irrespective of size or GC content, without bias
• The purified exosomal RNA is free from any circulating RNA-binding proteins
• No phenol extractions, Proteinase K treatment nor carrier RNA required
• No time-consuming ultracentrifugation, filtration nor special syringes are required
• No precipitation reagents nor overnight incubation required
• Concentrate isolated exosomal RNA and free-circulating RNA into a flexible elution volume ranging from 50 µL to 100 µL
• Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix

These kits provide a fast, reliable and convenient method to sequentially isolate and concentrate exosomal RNA as well as Free-Circulating RNA from different urine sample volumes. The purification is based on spin column chromatography that employs Norgen’s proprietary resin. These kits are designed to isolate all sizes of RNA, including microRNA, irrespective of size or GC content, without bias. These kits provide a clear advantage over other available kits since they do not require any special instrumentation, protein precipitation reagents, extension tubes, phenol/chloroform or protease treatments. Moreover, the kits allow the user to elute into a flexible elution volume ranging from 50 µL to 100 µL. The RNA isolated from the purified exosomes is free from any protein-bound circulating RNA and is of the highest integrity. Moreover, the free-circulating, protein-bound, RNA is free from any exosomal RNA. The purified RNA 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.

Urine Exosome and Free-Circulating RNA Isolation Mini Kit

For sample volumes ranging from 250 µL to 1 mL.

Urine Exosome and Free-Circulating RNA Isolation Midi Kit

For sample volumes ranging from 2 mL to 10 mL.

Urine Exosome and Free-Circulating RNA Isolation Maxi Kit

For sample volumes ranging from 11 mL to 30 mL.

Details

Supporting Data

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Kit Specifications
Minimum Urine Input	11 mL
Maximum Urine Input	30 mL
Size of RNA Purified	All sizes, including miRNA and small RNA (< 200 nt)
Elution Volume	50 – 100 µL
Time to Complete 10 Purifications	40 – 45 minutes
Average Yields*	Variable depending on the specimen

*Please check page 5 of the product insert for the average yields and the common RNA quantification methods.

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 shipping. It is recommended to warm Lysis Buffer A for 20 minutes at 60°C if any salt precipitation is observed.

Important Note
Urine samples stored at -70°C, -20°C or at 4°C will develop some precipitation due to the aggregation of some of the highly abundant proteins in urine. Eliminating these precipitates using centrifugation or filtration may cause the loss of exosomes. Furthermore, these precipitates may affect the quality of the purified nucleic acid. We recommend the use of Norgen’s Urine Preservative when collecting urine samples, which is designed for the preservation of nucleic acids and proteins in fresh urine samples at ambient temperatures. The components of the Urine Preservative allow samples to be stored for over 2 years at room temperature with no detected degradation of urine DNA, RNA or proteins. Norgen’s Urine Preservative is available as a liquid format in Norgen’s Urine Preservative Single Dose Ampules, as well as in a dried format in Norgen’s Urine Collection and Preservation Tubes.

Component	Cat. 59200 (50 preps)	Cat. 59300 (25 preps)	Cat. 59400 (15 preps)
Slurry E	12.5 mL	12.5 mL	12.5 mL
ExoC Buffer	8 mL	30 mL	50 mL
ExoR Buffer	12 mL	12 mL	12 mL
Lysis Buffer A	2 x 20 mL	2 x 20 mL	2 x 20 mL
Lysis Additive B	2 mL	2 mL	2 mL
Wash Solution A	2 x 18 mL	18 mL	18 mL
Elution Solution A	2 x 6 mL	6 mL	6 mL
Mini Filter Spin Columns	50	25	15
Mini Spin Columns	100	50	30
Collection Tubes	100	50	30
Elution Tubes (1.7 mL)	100	50	30
Product Insert	1	1	1

The NGS DNA Fragmentation & Library Prep Kit (illumina and MGI Platforms) was developed for construction of high quality libraries for next generation sequencing. The kit uses intact genomic DNA (EDTA-free DNA or DNA resuspended in TE buffer) as input DNA without an additional DNA fragmentation step. Our technology provides a fast and simple workflow. DNA libraries can be generated around 2 hours with only 10 min hands-on time. Library multiplexing is possible.

NGS DNA Fragmentation & Library Prep Kit Workflow

The incorporation of DNA fragmentation in the kit makes it possible to directly use intact genomic DNA as input DNA without the need of mechanical DNA shearing or enzymatic DNA fragmentation. The NGS DNA Fragmentation & Library Prep Kit does not generate sequencing bias as compared to library using mechanical sheared DNA as input. Sequence coverage is also consistent between enzymatic shearing and mechanical shearing. The library size is inversely correlated with the incubation time of step 1 at 20°C.

Three index types are available for the kit of the illumina platform:

Non-index (Cat.# 30026): Libraries do not have index.

Index (Cat.# 30028): Each of the index primers contains a unique index sequence of 6 bases. Library multiplexing for 48 samples is possible. Index information can be downloaded here.

Unique dual index (Cat.# 30030): Library multiplexing for 96 samples is possible. With the unique features of our 4-Base Difference Index System, the index sequence is 8-base long and each index has 4 bases different from others. Our unique dual index primers effectively identify sequencing errors such as index hopping, mis-assignment of reads, and de-multiplexing errors etc. The unique dual index primers set consists of 96 pre-mixed unique pairs of  index primers. Index information can be downloaded here.

Indexes are available for the MGI platform kits (Cat.# 34028).

Kit features:

• • • 1.5-hour protocol from intact genomic DNA to NGS library
    • Intact genomic DNA as input, DNA fragmentation is not needed.
    • Works with both EDTA-free DNA and DNA resuspended in TE buffer
    • Simple workflow: Less steps
    • Guaranteed quality: Higher library conversion efficiency

Library conversion efficiency: 100 ng, 300 ng and 500 ng of intact genomic DNA were used as input.

The library size is inversely correlated with the incubation time of step 1 at 20°C.

NGS data comparison: enzymatic shearing versus mechanical shearing

Enzymatic shearing
• DNA shearing and library prep: BioDynami NGS DNA Fragmentation & Library Prep Kit
Mechanical shearing
• DNA shearing: Covaris sonication
• Library prep: BioDynami NGS DNA Library Prep Kit.

The NGS DNA Fragmentation & Library Prep Kit (illumina and MGI Platforms) was developed for construction of high quality libraries for next generation sequencing. The kit uses intact genomic DNA (EDTA-free DNA or DNA resuspended in TE buffer) as input DNA without an additional DNA fragmentation step. Our technology provides a fast and simple workflow. DNA libraries can be generated around 2 hours with only 10 min hands-on time. Library multiplexing is possible.