Biotin-PEG8-DBCO

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

Overview

• Isolate and extract high quality & quantity miRNA in 30 minutes
• Two column kit- eliminate large RNA on the first column and capture microRNA on a second column
• Elute microRNA in 25 µL ready for miRNA profiling
• Process a wide range of samples- cell, tissue, bacteria, bodily fluids, etc.
• Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix

This kit provides a rapid method for the isolation and purification of small RNA molecules (< 200 nt) from cultured animal cells, small tissue samples, bacterial cells, plants, and blood.

Two columns are provided with this kit. The first column captures large RNA, while the small RNA are captured on a second column and are eluted concentrated in 25 µL of nuclease-free water.

The small RNA can be used in various downstream applications relating to miRNA profiling, gene regulation, and functional analysis. The eluted RNA is ready for RT-qPCR, microarrays, and NGS applications.

Background

These small RNAs include regulatory RNA molecules such as microRNA (miRNA) and short interfering RNA (siRNA), as well as tRNA and 5S rRNA. Small RNA molecules are often studied due to their ability to regulate gene expression. miRNAs and siRNAs are typically 20-25 nucleotides long and regulate gene expression by binding to mRNA molecules and affecting their stability or translation.

Details

Supporting Data

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Kit Specifications
Maximum Column Binding Capacity	Up to 50 μg RNA
Maximum Column Loading Volume	650 μL
Minimum Elution Volume	20 μL
Size of RNA Purified	All sizes, including small RNA < 200 nt
Time to Complete 10 Purifications	25 minutes
Maximum Amount of Starting Material: Animal Cells Animal Tissues Bacteria Plant Tissues Blood	3 x 106 cells 5-25 mg 1 x 109 cells 50 mg 100 μL

 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. 21300 (25 preps)
Buffer RL	40 mL
Wash Solution A	38 mL
Elution Solution A	6 mL
Large RNA Removal Column	25
microRNA Enrichment Column	25
Collection Tube	50
Elution Tubes (1.7 mL)	50
Product Insert	1

Propargyl-PEG5-CH2CO2tBu enables the formation of triazole linkages with azide compounds or biomolecules in copper catalyzed Click Chemistry reactions. The t-butyl group can be hydrolyzed under acidic conditions. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

Propargyl-PEG5-CH2CO2tBu enables the formation of triazole linkages with azide compounds or biomolecules in copper catalyzed Click Chemistry reactions. The t-butyl group can be hydrolyzed under acidic conditions. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.