Introduction

This product provides fast and easy methods for purification of total DNA for reliable PCR and Southern blotting. Total DNA(e.g., genomic, viral, mitochondrial) can be purified from small volume of blood, tissue and dry blood spots.

Details

Specifications

Features	Specifications
Main Functions	Isolation total DNA from 1-10μl blood, <10mg tissue, urine, blood stain, seminal stain
Applications	PCR, southern bolt and virus detection, etc.
Purification method	Mini spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	Animal tissues, blood stain, urine, seminal stain and various forensic samples
Sample amount	Blood:1-100μl, Tissue:<10mg
Elution volume
Time per run
Liquid carrying volume per column
Binding yield of column

Principle

This product is based on silica Column purification. The sample is lysed and digested with lysate and protease, DNA is released into the lysate. Transfer to an adsorption column. Nucleic acid is adsorbed on the membrane, while protein is not adsorbed and is removed with filtration. After washing proteins and other impurities, nucleic acid was finally eluted with low-salt buffer (10mmTris, pH9.0, 0.5mm EDTA).

Advantages

• Fast – several samples can be extracted in 20 minutes (after digestion)
• High purity – purified DNA can be directly used in various downstream applications
• High recovery – DNA can be recovered at the level of PG
• Good repeatability – silica technology can obtain ideal results every time

Kit Contents

Contents	D312502	D312503
Purification Times	50 Preps	250 Preps
Buffer ATL	15 ml	60 ml
Buffer AL	15 ml	60 ml
Buffer GW1*	22 ml	66 ml
Buffer GW2*	20 ml	2 x 50 ml
Carrier RNA	310 μg	2 x 310 µg
Proteinase K	24 mg	120 mg
Protease Dissolve Buffer	1.8 ml	10 ml
Buffer AE	15 ml	60 ml
HiPure DNA Mini Columns I	50	2 x 125
2 ml Collection Tubes	100	5 x 100

Storage and Stability

Carrier RNA and Proteinase K should be stored at 2–8°C upon arrival. However, short-term storage (up to 12 weeks) at room temperature (15–25°C) does not affect its performance. The remaining kit components can be stored dry at room temperature (15–25°C) and are stable for at least 18 months under theseconditions.

Experiment Data

This product provides fast and easy methods for purification of total DNA for reliable PCR and Southern blotting. Total DNA(e.g., genomic, viral, mitochondrial) can be purified from small volume of blood, tissue and dry blood spots.

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 V2-V3 Library Preparation Kit for Illumina consists of the reagents and components required for library preparation of the 16S V2-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 V2-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 V2-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 V2-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. 70300 (24 preps)	Cat. 70310, 70320, 70330, 70340 (96 preps)
Amplicon PCR (PCR 1)	MGX Master Mix	330 µL	1,320 µL
	16S V2-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 kit is suitable for extracting total pathogen nucleic acid from a variety of clinical samples (including serum and plasma). The kit is based on super paramagnetic particles purification technology. Purified DNA/RNA is ready for downstream applications such as Real Time PCR, biochip analysis, NGS and other related experiments.

Details

Specifications

Features	Specifications
Main Functions	Extract Pathogen RNA/DNA from 0.5ml plasma, serum, body fluid, homogenate suspension, culture solution, cell suspension, soaking solution or concentrate pathogen solution for mNGS application, remove host background nucleic acid.
Applications	RT-PCR，Real Time PCR, biochip analysis, NGS
Products	Pathogen DNA / RNA
Purification method	Polydisperse magnetic beads
Purification technology	Magnetic beads technology
Process method	Manual or automatic
Sample type	low/cell-free samples such as plasma, serum, body fluid, homogenate suspension, culture solution, cell suspension, soaking solution or concentrate pathogen solution
Sample amount	0.5 ml
Adaptive instrument	Nucleic acid extractor, pipetting workstation

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. After adding magnetic particles and binding solution, DNA/RNA 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/RNA was eluted by Buffer NFW.

Kit Contents

Contents	R667200B	R6672-02B
Purification Times	24 Preps	96 Preps
2ml Bead Tubes	24	96
Proteinase K	12 mg	50 mg
Protease Dissolve Buffer	1.8 ml	3 ml
Buffer SDS (20%)	1.8 ml	8 ml
MagBind Particles	0.6 ml	2.5 ml
Buffer MLB	15 ml	60 ml
Buffer MW1*	13 ml	44 ml
Buffer MW2*	6 ml	50 ml
Buffer AVE	5 ml	30 ml

Storage and Stability

MagBind Particles and Proteinase K Solution should be stored at 2–8°C upon arrival. However, short-term storage (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 18 months under these conditions.

This kit is suitable for extracting total pathogen nucleic acid from a variety of clinical samples (including serum and plasma). The kit is based on super paramagnetic particles purification technology. Purified DNA/RNA is ready for downstream applications such as Real Time PCR, biochip analysis, NGS and other related experiments.