Propargyl-PEG4-Maleimide has a propargyl group and a maleimide group. The propargyl group reacts with azide compounds or biomolecules under the catalyzation of copper in Click Chemistry reactions. The maleimide group enables the formation of a covalent bond with biomolecules containing a thiol group. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

Propargyl-PEG4-Maleimide has a propargyl group and a maleimide group. The propargyl group reacts with azide compounds or biomolecules under the catalyzation of copper in Click Chemistry reactions. The maleimide group enables the formation of a covalent bond with biomolecules containing a thiol group. 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 V3-V5 Library Preparation Kit for Illumina consists of the reagents and components required for library preparation of the 16S V3-V4 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 V3-V5 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 V3-V4 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

Figure 1 / 3

Previous

Next

Click for expanded view

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 V3-V5Library 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. 70500 (24 preps)	Cat. 70510, 70520, 70530, 70540 (96 preps)
Amplicon PCR (PCR 1)	MGX Master Mix	330 µL	1,320 µL
	16S V3-V5 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

A rapid, high performance dye-terminator removal process based on the paramagnetic bead technology. The paramagnetic bead format requires no centrifugation or filtration and is easily performedmanually or fully automated for high throughput dye-terminator removal. Compared to similar systems, this product produces sequences with longer Phred 20 read lengths and higher signal intensities than any other purification technology.

Details

Specifications

Features	Specifications
Main Functions	Removal of free fluorescent dye from sequencingsolution (Replace Beckmen or agencourt CleanSeq)
Applications	Automated sequences
Purification technology	Magnetic beads technology
Process method	Manual or automatic
Sample type	DNA doped with fluorescentdye
Sample amount	5μl
Recovery	90%
Elution volume	≥25μl
Operation time	≤50 minutes

Principle

The CleanSeq method contains magnetic particles in an optimized binding buffer to selectively capture sequencing extension products. The protocol can be performed directly in the thermal cycling plate. Unincorporated dyes, nucleotides, salts and contaminants are removed using a simple washing procedure. The purification procedure is amenable to a variety of automation platforms since it requires no centrifugation or vacuum filtration.

Advantages

• High recovery – up to 90%
• High throughput – using magnetic beads purification technology
• Simple – simplified operation process, combination – washing – elution

Kit Contents

Contents	BCS-5	BCS-50	BCS-500
CleanSeq Beads	5 ml	50 ml	500 ml

Storage and Stability

CleanSeq Beads should be stored at 2-8°C upon arrival and is stable up to 6 months under the condition. However, short-term storage (up to 12 weeks) at room temperature (15-25°C) does not affect its performance. Mix CleanSeq Beads well before using. The reagent should appear homogenous and consistent in color.

DO NOT FREEZE.

A rapid, high performance dye-terminator removal process based on the paramagnetic bead technology. The paramagnetic bead format requires no centrifugation or filtration and is easily performedmanually or fully automated for high throughput dye-terminator removal. Compared to similar systems, this product produces sequences with longer Phred 20 read lengths and higher signal intensities than any other purification technology.