Overview

• Protocol optimized for DNA isolated from a diversity of samples including stool, soil, water, saliva, plant, urine, skin, and more
• Simple and quick workflow: library could be prepared in less than 4 hours
• Component of Norgen’s metagenomics workflow
• A single NGS run can be prepared with up to 384 unique dual-index libraries

Sample type purification kit guide 

The ITS2 Library Preparation Kit for Illumina consists of the reagents and components required for library preparation of the fungal target ITS2 libraries to be used for next-generation sequencing on Illumina platforms. All molecular reagents including primers, enzyme mixes, indexes, and buffers are provided. Instructions for PCR clean up with the AMPure XPMagneticBeads(supplied by customer) are also included for rapid purification of nucleic acid products generated at two steps of the workflow. The library prep workflow could be used for purified DNA inputs from different sources including stool, soil, water, saliva, plant, urine, skin swab, vaginal swab, cheek swab, nasal swab, plasma/serum, tongue swab, gum swab, and others.

The ITS2 Library Preparation Kit for Illumina has a streamlined procedure that reduces the handling time such that the library prep procedure can be completed in approximately 4 hours (see diagram below). Input DNA is first subjected to targeted PCR to amplify the ITS2 region of the fungal DNA. The post-PCR reaction is then cleaned up using AMPure XPbeads. Dual index primers are then added using a limited-cycle PCR. The indexed amplicons flanked by 5′ and 3′ barcoded adaptors are then cleaned using AMPure XPbeads. The libraries are then ready for quantification, pooling and sequencing.

Details

Supporting Data

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Storage Conditions and Product Stability
Norgen’s ITS2 Library Prep Kit for Illumina is shipped as one kit box (for the 24 prep kit) or two sub-component kits (for the 96 prep kit). All kits should be stored at -20°C upon arrival.

All kit components will remain stable for at least 1 year when stored at the specified storage conditions.

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

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

Purchase Guide

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.

Introduction

This kit provides a simple and fast solution for the extraction of circulating nuclear acid from serum, plasma, and other cell-free liquid samples. Circulating nucleic acid refers to the free extracellular nucleic acid produced by cell apoptosis, of which fragments are generally below 1KB. The kit is based on silica gel column purification technology, which is no need for toxic phenol chloroform extraction and time-consuming alcohol precipitation during the extraction. The obtained Circulating Nucleic Acid can be directly used for quantitative PCR, liquid or solid phase chip analysis, hybridization, and SNP detection.
HiPure Circulating DNA/RNA Kit adopts a unique solution system and multiple layers of filter membranes with different pore sizes, which can efficiently process large volumes of serum and plasma samples and capture extremely small amounts of free nucleic acids.

Details

Specifications

Principle

This kit is based on silica gel column technology. Serum or other liquid samples are lysed and digested in buffer CFL. After adding buffer CFP, the protein is removed by centrifugation to obtain the supernatant. Isopropanol is added to precipitate the total nucleic acid and transferred to the column for filtration. DNA / RNA is adsorbed on the membrane of the column, while the protein is not adsorbed and removed with the filtrate.The column is washed with buffer MGW1 to remove protein and other impurities, and then washed with buffer RW2 to remove salt. Finally, DNA / RNA is eluted by low salt buffer. The eluted DNA / RNA can be directly used for quantitative PCR/ RT-PCR, liquid or solid-phase chip analysis, hybridization and SNP detection.

Advantages

• High yield – most optimized process to obtain maximum free RNA and small RNA
• High concentration – low elution volume (>20μl) to ensure nucleic acid concentration
• High purity – low alcohol combination, completely remove inhibitor and protein pollution
• High recovery – silica gel column technology can recover nucleic acid molecules at the level of PG
• Large volume – 1-2ml serum and plasma samples can be processed at one time

Kit Contents

Storage and Stability

The kit components can be stored at room temperature (15–25°C) and are stable for at least 18 months under these conditions.

This kit provides a simple and fast solution for the extraction of circulating nuclear acid from serum, plasma, and other cell-free liquid samples. Circulating nucleic acid refers to the free extracellular nucleic acid produced by cell apoptosis, of which fragments are generally below 1KB. The kit is based on silica gel column purification technology, which is no need for toxic phenol chloroform extraction and time-consuming alcohol precipitation during the extraction. The obtained Circulating Nucleic Acid can be directly used for quantitative PCR, liquid or solid phase chip analysis, hybridization, and SNP detection.
HiPure Circulating DNA/RNA Kit adopts a unique solution system and multiple layers of filter membranes with different pore sizes, which can efficiently process large volumes of serum and plasma samples and capture extremely small amounts of free nucleic acids.