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-V3 Library Preparation Kit for Illumina consists of the reagents and components required for library preparation of the 16S V1-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 V1-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 V1-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 V1-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. 70200 (24 preps)	Cat. 70210, 70220, 70230, 70240 (96 preps)
Amplicon PCR (PCR 1)	MGX Master Mix	330 µL	1,320 µL
	16S V1-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

Introduction

This product supplies a simple and rapid extraction of total RNA including microRNA from Blood, buffy coat, bone marrow, Cell suspension and other body fluids. The kit is based on superparamagnetic particles purification technology, no phenol-chloroform extraction or alcohol precipitation. Purified RNA is ready for downstream applications such as RT-PCR, virus RNA testing and so on.

Details

Principle

The Kit can be used for both manual extraction process and automatic nucleic acid extraction machines. This Kits is suitable for extracting RNA from ≤ 1 x10⁶ cells suspension, 50 μl Whole Blood, 50μl buffy coat, 20μl bone marrow. This product is based on the purification method of high binding magnetic particles. The sample is lysed and digested under the action of lysate and Protease. RNA/DNA is released into the lysate. After adding magnetic particles and binding solution, DNA/RNA will be adsorbed on the surface of magnetic particles, and impurities such as proteins will be removed without adsorption. The adsorbed particles were digested using DNase and washed with washing buffer to remove proteins and impurities, washed with ethanol to remove salts, and

Kit Contents

Contents	R661101C	R661102C	R661103C
Purification times	48 Preps	96 Preps	480 preps
MagPure RNA Particles	1.0 ml	1.5 ml	6 ml
Proteinase K	24 mg	48 mg	220 mg
Protease Dissolve Buffer	1.8 ml	1.8 ml	15 ml
DNase I	600 μl	2 x 600 μl	10 x 600 µl
DNase Buffer	15 ml	15 ml	60 ml
Buffer AL	10 ml	10 ml	60 ml
Buffer MCB*	9 ml	9 ml	30 ml
Buffer MW1*	13 ml	22 ml	110 ml
Buffer MW2*	6 ml	20 ml	100 ml
RNase Free Water	10 ml	15 ml	120 ml

Storage and Stability

MagPure RNA Particles and 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, MagPure RNA Particles and 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 atleast 18 months under these conditions.

This product supplies a simple and rapid extraction of total RNA including microRNA from Blood, buffy coat, bone marrow, Cell suspension and other body fluids. The kit is based on superparamagnetic particles purification technology, no phenol-chloroform extraction or alcohol precipitation. Purified RNA is ready for downstream applications such as RT-PCR, virus RNA testing and so on.

Overview

• Fast and easy recovery of DNA from agarose gel fragments
• High recovery of desired DNA
• Convenient spin column format
• DNA is ready for ligation, restriction digestion, sequencing and more

This kit is designed for the rapid preparation and purification of DNA fragments that have been fractionated on agarose gels. The recovered DNA is free from agarose and other impurities, and is compatible with restriction enzyme digestion, ligation into vectors and sequencing.  The protocol can be completed in 20 minutes.  

Details

Supporting Data

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Kit Specifications
Column Binding Capacity	10 μg
Maximum Weight of Gel Slice	400 mg
Average Recovery	70-90%
Size of DNA Purified	100 – 15,000 bp
Minimum Elution Volume	30 μL
Time to Complete 10 Purifications	30 minutes

Storage Conditions and Product Stability

All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 1 year after the date of shipment.

Component	Cat. 13100 (50 preps)
Binding Buffer G	80 mL
Wash Solution A	12 mL
Elution Buffer B	8 mL
Spin Columns	50
Collection Tubes	50
Elution Tubes (1.7 mL)	50
Product Insert	1