Sulfo DBCO-PEG3-NHS ester is a water soluble, click chemistry reagent which can undergo copper free click chemistry Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

Sulfo DBCO-PEG3-NHS ester is a water soluble, click chemistry reagent which can undergo copper free click chemistry Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

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 is suitable for rapid extraction of DNA from tissue, cells, blood, saliva, swabs, blood spots, semen and other clinical samples. DNA can be used directly for PCR, quantitative PCR, Southern Blot, test of virus DNA and so on.

Details

Specifications

Features	Specifications
Main Functions	Isolation total DNA from blood, buffy coat, tissue and other samples
Applications	Second generation sequencing, PCR, real time PCR, etc.
Purification method	Polydisperse magnetic beads
Purification technology	Magnetic beads technology
Process method	Manual or automatic
Sample type	Anticoagulant blood, concentrated blood, buffy coat, lymphocytes and cultured cells
Sample amount	Whole blood :< 200μl; Saliva / swab:< 400μl; Tissue :< 20mg
Yield	0.1 – 50μg
Elution volume
Time per run

Principle

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. DNA is released into the lysate. After adding magnetic particles and binding solution, DNA will be adsorbed on the surface of magnetic particles, and impurities such as proteins will be removed without adsorption. The adsorbed particles were washed with washing solution to remove proteins and impurities, washed with ethanol to remove salts, and finally DNA was eluted by Elution Buffer.

Advantages

• High purity – OD 260 / 280 : 1.7- 1.9, OD 260 / 230 : 1.5 – 2.0
• Economy – less than 50% of the price of Qiagen and other imported products
• High yield – most optimal process, ensuring the recovery up to 90%
• Strong processing ability – samples including animal blood, cultured cells, animal tissues, etc.

Kit Contents

Contents	IVD3102
Purification Times	200
MagPure Particles	5 ml
Proteinase K	100 mg
Protease Dissolve Buffer	10 ml
Rnase A	40 mg
Buffer ATL	60 ml
Buffer AL	60 ml
Buffer BD*	20 ml
Buffer BW1*	110 ml
Elution Buffer	30 ml

Cat.No	Reagent	IVD3102-F-96
Proteinase K		50 mg
Protease Dissolve Buffer		6 ml
RNase A		20 mg
Buffer ATL		30 ml
Buffer AL		30 ml
96-Tip		1
Sample plate (DW Plate)	450µl Buffer BD(Ethanol Added)	1
Wash 1 Plate (DW Plate)	600µl Buffer BW1(Ethanol Added)	1
Wash 2 Plate (DW Plate)	600µl Buffer BW1(Ethanol Added)	1
Wash 3 Plate (DW Plate)	750µl Buffer GW2, 20µl MagPure Particle	1
Elution plate (DW Plate)	80µl Elution Buffer	1

Cat.No	Reagent	IVD3102-TL-06
Proteinase K		50 mg
RNase A		20 mg
Protease Dissolve Buffer		6 ml
Buffer ATL		40 ml
Buffer AL		40 ml
AS-Tip		12
2.0ml V-bottom plate	Row 1/7：450µl Buffer BDRow 2/8：450µl Buffer BW1Row 3/9：450µl Buffer BW1Row 4/10：20μl Magpure Particle450μl Wash Buffer GW2 Row 5/11：450μl Wash Buffer GW2 Row 6/12：80µl Elution Buffer	6

Storage and Stability

Proteinase K, RNase A, MagPure Particles should be stored at 2-8°C upon arrival. However, short-term storage (up to 24 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 at least 18 months under these conditions.

Experiment Data

This product is suitable for rapid extraction of DNA from tissue, cells, blood, saliva, swabs, blood spots, semen and other clinical samples. DNA can be used directly for PCR, quantitative PCR, Southern Blot, test of virus DNA and so on.