Introduction

This kit is designed to rich and extract 100bp-500bp circulating DNA from 5 ml cell-free body fluids (such asplasma, serum), and remove fragments above 500bp. Machine reaction only takes 90 minutes. Magnetic-particle technology provides high-quality DNA that is suitable for direct use in downstream applications such as PCR and next generation sequencing.

Details

Specifications

Features	Specifications
Main Functions	Isolation circulating DNA from 5ml plasma, serum, body fluids
Applications	qPCR, NGS, etc.
Purification technology	Magnetic beads technology
Process method	Manual (centrifugation or vacuum)
Sample type	Serum, plasma
Sample amount	5ml
Elution volume	≥40μl
Time per run	≤50 minutes

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

• Economy – less than 50% of the price of Qiagen and other imported products
• Automatic – without labour

Kit Contents

Contents	1292750	12927200
Purification Times	50	200
MagPure Particles G	20 ml	80 ml
MagBind Particles (selection particles)	14 ml	58 ml
Selection Solution	100 ml	400 ml
Proteinase K	300 mg	1.2 g
Protease Dissolve Buffer	25 ml	100 ml
Buffer SDS(20%)	15 ml	60 ml
Buffer MLK	300 ml	3 x 450 ml
Buffer BST1	225 ml	2x 450 ml
Buffer MKW1	225 ml	2x 450 ml
Buffer MW2*	50 ml	2x 100 ml
Buffer AE	10 ml	30 ml

Storage and Stability

MagPure Particles G, MagBind Particles and Proteinase K should bestored at 2–8°C upon arrival. However, short-term storage (up to 12 weeks) at room temperature (15–25°C) does not affect their performance. The remaining kit components can be stored dry at roomtemperature (15–25°C) and are stable for at least 18 months underthese conditions.The entire kit can be stored at 2–8°C, but in this case buffers should be redissolved before use. Make sure that all buffers are at room temperature when used.

This kit is designed to rich and extract 100bp-500bp circulating DNA from 5 ml cell-free body fluids (such asplasma, serum), and remove fragments above 500bp. Machine reaction only takes 90 minutes. Magnetic-particle technology provides high-quality DNA that is suitable for direct use in downstream applications such as PCR and next generation sequencing.

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-V2 Library Preparation Kit for Illumina consists of the reagents and components required for library preparation of the 16S V1-V2 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-V2 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-V2 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-V2 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. 70100 (24 preps)	Cat. 70110, 70120, 70130, 70140 (96 preps)
Amplicon PCR (PCR 1)	MGX Master Mix	330 µL	1,320 µL
	16S V1-V2 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

We will help to establish your experiments using our broad knowledge:

• With myPOLS Biotec’s access to large libraries of different mutant polymerases and experience in high throughput screening we can find the best enzyme for your needs.
• We also optimize your polymerases for your defined requirements or enhance our DNA polymerase-based products accordingly.
• Finally, we can develop freeze-dried enzyme formulations and production.

We at myPOLS Biotec are scientists and have vast experience in polymerase-based product development. You can benefit from this. Get in touch.

We will help to establish your experiments using our broad knowledge:

With myPOLS Biotec’s access to large libraries of different mutant polymerases and experience in high throughput screening we can find the best enzyme for your needs.
We also optimize your polymerases for your defined requirements or enhance our DNA polymerase-based products accordingly.
Finally, we can develop freeze-dried enzyme formulations and production.