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 V2-V3 Library Preparation Kit for Illumina consists of the reagents and components required for library preparation of the 16S V2-V3 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 V2-V3 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 V2-V3 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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Minimum amount of starting material:	2.5 µL of DNA (5 ng/µL)
Time to complete library preparation:	4 hours

Storage Conditions and Product Stability
Norgen’s 16S V2-V3 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.

Step	Component	Cat. 70300 (24 preps)	Cat. 70310, 70320, 70330, 70340 (96 preps)
Amplicon PCR (PCR 1)	MGX Master Mix	330 µL	1,320 µL
	16S V2-V3 Primer Mix	70 µL	280 µL
Index PCR (PCR 2)	Indexing Master Mix	660 µL	2 x 1,320 µL
	N7xx Index Primer	50 µL	50 µL
	S5xx Index Primer	70 µL	70 µL
PCR Clean-Up	Resuspension Buffer	2 x 1,250 µL	2 x 5,000 µL
	Nuclease-free water	1,250 µL	1 x 6,000 µL

Overview

• Detection kits for NoV
• CE-IVD marked version available for in vitro diagnostic use
• Available in TaqMan format for analysis

Norovirus is a single-stranded, non-enveloped RNA virus belonging to the Caliciviridae family. Norovirus is considered the major causative agent of non-bacterial gastroenteritis in the world. The main channel of transmittance is via contaminated food or water, as well as human-to-human contact. Most Noroviruses infecting humans belong to genogroup I and II (particularly genotype 4). Norovirus is very stable in the environment and is resistant to some surface disinfectants such as alcohols and detergents. Moreover, it is very difficult to culture Norovirus in vitro, making its identification by standard microbiological assays challenging.

NoV TaqMan RT-PCR Kit, 100 reactions

• Ready to use format, including Master Mix for the target and PCR control to monitor for PCR inhibition and validate the quality
• Specific Primer and Probe mix for the pathogen/virus/viroid of interest
• Primer and Probe mix
• Positive and negative control to confirm the integrity of the kit reagents

NoV TaqMan RT-PCR Probe/Primer Set and Controls, 100 reactions

• Specific Primer/Probe mix and Positive Control for the pathogen/virus/viroid of interest
• Nuclease-free water
• Can be used together with Norgen’s RT-PCR Master Mix (#28113) or customer supplied master mix

Details

Supporting Data

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Storage Conditions and Product Stability
All kit components can be stored for 1 year after the date of production without showing any reduction in performance.

All kit components should be stored at -20°C upon arrival.

Component	Cat. TM41450 (100 preps)	Cat. TM41410 (100 preps)
MDx TaqMan 2X PCR Master Mix	2 x 700 μL	–
NoV Primer & Probe Mix	280 μL	280 μL
NoV Positive Control	150 μL	150 μL
Nuclease-Free Water (Negative Control)	1.25 mL	1.25 mL
Product Insert	1	1

Introduction

HiPure DNA Clean Up 96 Kit uses proprietary chemistry and HiPure technology to recover DNA Fragments between 60bp-10kbp with yields exceeding 80%. DNA is suitable for ligations, PCR, sequencing, restriction digestion, or various labeling reactions. In addition, this kit can be also used to recover DNA directly from enzymatic reactions such as PCR and enzyme digestion reactions.

Details

Specifications

Features	Specifications
Main Functions	Recover DNA fragments between 60bp-10kbp from PCR, Reactions, crude DNA using 96 well Bind Plate
Applications	PCR, sequencing, labeling reactions, ligations and restriction digestion, etc.
Purification method	96 well Bind Plate
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	PCR product, enzymatic reaction
Sample amount	Appropriate
Recovery	≥80%
Elution volume	≥75μl
Time per run	≤60 minutes
Liquid carrying volume per column	800µl
Binding yield of column	20µg

Principle

The HiPure system uses a simple bind-wash-elute procedure. Gel slices are dissolved in a buffer containing a pH indicator, allowing easy determination of the optimal pH for DNA binding, and the mixture is applied to the column. Nucleic acids adsorb to the silica-gel membrane in the high-salt conditions provided by the buffer. Impurities are washed away and pure DNA is eluted with a small volume of low-salt buffer provided or water, ready to use in subsequent applications.

Advantages

• High recovery efficiency – ≥80% DNA recovery
• Fast – isolation can be completed in 60 minutes

Kit Contents

Contents	D212201	D212202	D212203
Purification Times	1 x 96 Preps	4 x 96 Preps	20 x 96 Preps
Buffer GDP	120 ml	400 ml	4 x 400 ml
Buffer DW2	20 ml	100 ml	3 x 100 ml
Elution Buffer	20 ml	20 ml	30 ml
HiPure DNA Plate	1	4	20
2.2ml Collection Plate	1	4	20
1.6 ml Collection Plate	1	4	20
0.8 ml Collection Plate	1	4	20

Storage and Stability

The Kit can be stored dry at room temperature (15-25°C) and are stable for at least 18 months under these conditions. If any precipitates form in the buffers, warm at 37℃ to dissolve.

HiPure DNA Clean Up 96 Kit uses proprietary chemistry and HiPure technology to recover DNA Fragments between 60bp-10kbp with yields exceeding 80%. DNA is suitable for ligations, PCR, sequencing, restriction digestion, or various labeling reactions. In addition, this kit can be also used to recover DNA directly from enzymatic reactions such as PCR and enzyme digestion reactions.