endo-BCN-PEG3-amine is a click chemistry linker containing a BCN and an amine group. The BCN group can react with azide-tagged biomolecules under mile condition. The amino group is reactive with carboxylic acids, activated NHS esters, carbonyls (ketone, aldehyde) etc. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

endo-BCN-PEG3-amine is a click chemistry linker containing a BCN and an amine group. The BCN group can react with azide-tagged biomolecules under mile condition. The amino group is reactive with carboxylic acids, activated NHS esters, carbonyls (ketone, aldehyde) etc. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

Introduction

This product is suitable for rapid extraction of RNA (include miRNA) from tissue, cells, blood, s and other clinical samples. RNA can be used directly for RT-PCR, quantitative RT-PCR, test of virus DNA and so on.

Details

Specifications

Features	Specifications
Main Functions	Isolation total RNA（miRNA）from tissue, cell using two columns and DNase plus reagent
Applications	RT-PCR, cDNA synthesis, second generation sequencing
Products	RNA, miRNA
Purification method	Mini spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	Clinical tissues, cells, lymphocytes
Sample amount	Tissue: <20 mgCells: <5 x 106
Yield	2-50μg
Elution volume	≥30μl
Time per run	≤25 minutes

Principle

This product is based on silica column purification. Remove paraffin by Buffer DPS. Sample lysis withproteinase K digestion requires only 15 minutes. After lysis, samples are incubated at 80ºC for 15 minutes. Transfer to an adsorption column and RNA is adsorbed on the membrane, while protein is not adsorbed andis removed with filtration. After washing proteins and other impurities, RNA was finally eluted with low-saltbuffer.

Advantages

• Efficient DNA removal – one step RNA extraction can effectively remove genomic DNA
• High quality – one step RNA extraction reagent combined with silica gel column can obtain the highest concentration
• Fast – the whole extraction only takes 15-25 minutes
• Nontoxic – no toxic phenol chloroform extraction is required in the extraction

Kit Contents

Contents	IVD4121
Purification Times	50 Preps
HiPure DNA Mini Column Ⅱ	50
HiPure RNA Mini Columns	50
2ml Collection Tubes	150
Proteinase K	24 mg
Protease Dissolve Buffer	1.8 ml
DNase I	600 μl
DNase Buffer	6 ml
Buffer RTL	40 ml
RNA Digestion Buffer	15 ml
Buffer RWC*	20 ml
Buffer RW2*	20 ml
Nuclease Free Water	10 ml

Storage and Stability

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, 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 at least 18 months under these conditions.

This product is suitable for rapid extraction of RNA (include miRNA) from tissue, cells, blood, s and other clinical samples. RNA can be used directly for RT-PCR, quantitative RT-PCR, test of virus DNA and so on.

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-V4 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-V4 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

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-V4 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. 70400 (24 preps)	Cat. 70410, 70420, 70430, 70440 (96 preps)
Amplicon PCR (PCR 1)	MGX Master Mix	330 µL	1,320 µL
	16S V3-V4 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