Introduction

Free-circulating nucleic acids, such as tumor-specific extracellular DNA fragments and mRNAs in the blood or fetal nucleic acids in maternal blood, are present in serum or plasmausually as short fragments, <1000bp(DNA). HiPure Circulating DNA Midi Kit enables efficient purification of these circulating nucleic acids from human plasma, serum, or urine. The extracted products can be used for clinical in vitro detection.

Details

Specifications

Principle

This product is based on silica gel purification. The sample is lysed and digested with lysate and protease, DNA is released into the lysate. Transfer to an adsorption plate and filter column. Nucleic acid is adsorbed on the membrane, while protein is not adsorbed and is removed with filtration. After washing proteins and other impurities, nucleic acid was finally eluted with low-salt buffer (10 Mm Tris,pH 8.0).

Advantages

• High yield – most optimal process, free DNA (>50bp) can be obtained to the maximum extent
• High concentration – low elution volume, ensuring high nucleic acid concentration
• High purity – low alcohol binding method, completely removing inhibitor and protein pollution
• High recovery – DNA can be recovered at thelevel of PG by silica gel column purification

Kit Contents

Storage and Stability

Proteinase K, Carrier RNA should be stored at 2-8°C upon arrival. However, short-term storage (up to 12weeks) at room temperature (15-25°C) does not affect their performance. The remaining kit components can be stored dry at room temperature (15-25°C) and are stable for at least 18 months under these conditions. The entire kit can bestored at 2-8°C, but in this case buffers should be redissolved before use. Make sure that all buffers are at room temperature when used.

Free-circulating nucleic acids, such as tumor-specific extracellular DNA fragments and mRNAs in the blood or fetal nucleic acids in maternal blood, are present in serum or plasmausually as short fragments,

Overview

• Convenient optimized on-column DNase treatment using Norgen’s RNA Purification Kits
• Also includes protocol for digestion in-solution followed by RNA Clean-Up
• Guaranteed RNase-Free
• Includes Enzyme Incubation Buffer
• Cat. 25710 contains one vial of 1,600 units and Cat. 25720 contains 4 vials (1,600 units/vial)

Norgen’s RNA purification kits isolate total RNA with minimal amounts of genomic DNA contamination. However, for some sensitive downstream applications, it may be desirable to remove all traces of residual DNA. Norgen’s RNase-free DNAse I Kit, with Enzyme Incubation Buffer, can be used for optional on-column DNase digestion with any of Norgen’s RNA purification kits. Alternatively, after isolating total RNA using one of Norgen’s RNA purification kits, the RNA elution can be treated with this DNase I. The RNA can then be purified from the DNase using Norgen’s RNA Clean-Up and Concentration Kit (Cat# 23600), and the RNA can then be used in downstream applications.

Details

Each RNase-Free DNase I Kit is supplied complete with sufficient enzyme and enzyme incubation buffer for 50 or 200 reactions.

Storage Conditions
The DNase I provided is in lyophilized form. It is stable for at least 3 months if stored at room temperature. However, it is recommended to store the DNase I vial at 2 – 8ºC (or below) upon receipt to maintain stability beyond 3 months. Buffer DR and Enzyme Incubation Buffer can be stored at room temperature. After reconstitution with Buffer DR (see product manual), the DNase I should be stored at -20ºC. All reagents should remain stable for at least 1 year in their unopened containers at the appropriate storage temperature.

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 5 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 16S V1-V2 Library Preparation Kit for Illumina consists of the reagents and components required for library preparation of the 16S V1-V2 amplicon 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 XP Magnetic Beads (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 16S V1-V2 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 V1-V2 region of the DNA encoding 16S rRNA. The post-PCR reaction is then cleaned up using AMPure XP beads. 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 XP beads. The libraries are then ready for quantification, pooling and sequencing.

Details

Supporting Data

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Storage Conditions and Product Stability
Norgen’s 16S V1-V2 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 should remain stable for at least 1 year when stored at the specified storage conditions.