4x4x15ml Swing Buckets Rotor 

Overview

• DNA can be isolated and detected from as little as 100 µL of sputum
• Effective removal of PCR inhibitors
• Compatible with Norgen’s Saliva Collection and Preservation Device (Cat. RU49000)
• Sputum Liquification Buffer (Cat. 28289) can be purchased separately

This kit constitutes an all-in-one system for the rapid isolation of DNA from sputum samples. It allows for the isolation of bacterial or eukaryotic DNA from the sputum samples using spin-column chromatography based on Norgen’s proprietary resin. The kit includes all the reagents needed to process sputum DNA. The protocol can be completed in 30 minutes. Purified DNA is of the highest quality and free from inhibitors, and can be used in sensitive downstream applications including PCR and qPCR.

Background

A sputum specimen is the name given to the mucus which is expectorated from the lower airways. High quality sputum samples should contain very little saliva, as this may contaminate the sputum sample with oral bacteria. Sputum samples are typically evaluated to look for infections such as Moraxella catarrhalis, Mycobacterium tuberculosis, Streptococcus pneumoniae ;and Haemophilus influenza. Other pathogens can be detected in sputum as well, including HIV. In addition, sputum samples are useful in the detection of lung cancer or to evaluate chronic inflammation.

Details

Supporting Data

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Kit Specifications
Time to Complete DNA Isolation	< 1 hour
Average Yield of DNA from 1.0 mL*	75 μg
Average Purity (OD260/280)	1.8 – 2.1

* Average DNA yield will vary depending on the health status of the donor

Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment.

Norgen’s Sputum DNA Isolation Kit contains ready-to-use Proteinase K, which is dissolved in a specially prepared storage buffer. The buffered Proteinase K is stable for up to 2 years after the date of shipment when stored at room temperature.

Component	Cat. 46200 (25 preps)
Slurry D	55 mL
Proteinase K in Storage Buffer	0.6 mL
Solution WN	4 mL
Wash Solution BE	9 mL
Elution Buffer B	8 mL
Mini Filter Spin Columns	25
Collection Tubes	25
Elution Tubes (1.7 mL)	25
Product Insert	1

Introduction

Magnetic bead nucleic acid purification technology uses nano or micron superparamagnetic material as the matrix, generally black ferric oxide or yellowish brown ferricoxide as the magnetic material. The surface of bead is coated with appropriate functional groups, which can adsorb nucleic acid. Magnetic beads commonly used for nucleic acids, containing carboxyl groups, hydroxyl groups, or silicon groups. Silicon-based magnetic beads are the most common, and its principle of adsorbing nucleic acid is consistent with the classical glass milk purification technology or glass fiber filter membrane purification method. Magpure particle is a kind of polydisperse fast speed silica magnetic beads. The core is ferricoxide, accounting for 50%, and the surface coating is silica, accounting for 50%. The product can be used for plasmid extraction, gel DNA recovery, product purification, genomic DNA and RNA extraction, and viral nucleic acid extraction.

Details

Specifications

Features	Specifications
Concentration	40 mg/ml
Appearance	Suspension of dark brown particles
Surface functional group	Si-OH, Silanol
Dispersibility	Monodisperse,spherical
Particle size	1.0-1.5 μm
Preservation conditions	Room Temperature, valid for up to 2 years.It is recommended to store in 2-8°C to prevent microbial growth.
Magnetic response speed	~30 seconds
Settling velocity	>3 minutes
High salt mediated binding	>2M guanidine isothiocyanate, DNA recovery up to 80%
Alcohol mediated binding	2M guanidine hydrochloride / isopropanol (30%), and the recovery of DNA / RNA was as high as 85%
PEG8000 mediated binding	The recovery of DNA/RNA was up to 85%
DNase/RNase	Not detected
DNA residue	Not detected
Recommended application	Genomic DNA extraction, RNA extraction, viral nucleic acid extraction, circulating DNA isolation

Principle

Highsalt mediated binding: in the solution containing 2-4M guanidine isothiocyanate, Magpure particles can selectively recover DNA molecules, and impurities such as protein polysaccharides are not adsorbed.

Alcohol mediated binding: in the solution containing guanidine salt and alcohol (>25%), Magpure particles can selectively recover DNA/RNA molecules, and proteins and other impurities are not adsorbed.

After biological samples are treated with digestive solution or lysis Buffer, DNA/RNA is released from cells, organelles and protein complexes (ribosomes and nucleosomes) into reagents. After Magpure particles and binding solution are added, DNA/RNA is adsorbed to the surface of Magpure particles to form DNA/RNA bead complex. Under the action of the magnetic field, the magnetic beads are separated and collected, and the impurities such as protein are removed with the waste liquid. After two or three steps of further cleaning, the DNA/RNA magnetic bead complex is resuspended in sterilized water or TE buffer, and the DNA/RNA falls off from the surface of the magnetic beads, so as to achieve the purpose of purification.

Ordering information

CAT.No.	Product Name	Package
C14120	MagPure Particles G	100 ml
C14121	MagPure Particles G	400 ml
C14122	MagPure Particles G	3 x 400 ml
C14123	MagPure Particles G	10 x 400 ml

Purchase Guide

Features	MagPure Particles	MagPure Particles N	MagPure Particles G	MagPure Particles F	MagBind Particles
Cat.No.	C1410	C1411	C1412	C1414	C1413
Concentration	100mg/ml	70mg/ml	40mg/ml	50mg/ml	10mg/ml
Form	Amorphous and Porous	Amorphous and Porous	Porous	Amorphous	Nonporous
Surface function	Si-OH, Silica Beads	Si-OH, Silica Beads	Si-OH, Silica Beads	Si-OH, Silica Beads	COOH, Carboxyl Beads
Dispersion	Polydisperse	Polydisperse	Monodisperse	Monodisperse	Monodisperse
Particle Size	1.5-5μm	0.2-2μm	1-1.5μm	0.2-1.5μm	0.8-1μm
Color	Black	Yellowish Brown	Dark Brown	Dark Brown	Yellowish Brown
Magnetic response	15-30s	~60s	~30s	20s	120s
Settling Time (1ml)	>5min	>10min	>3min	>3min	>2h
Usage (0.2ml Sample)	20μl	20μl	20-30μl	20-30μl	20-30μl
DNA Recover Rate (only 4M GITC)	>80%	>80%	>80%	>80%	0
DNA Recover Rate (10% PEG8000/NaCl)	>85%	>85%	>85%	>85%	>90%
Recommended Use	gDNA/RNA Isolation from Blood, Tissue, Plant, Swab, Spots, Stool, Soil and etc.Viral DNA/RNA IsolationAgarose Gel DNA Purification	DNA/RNA Isolation from low nucleic acid content samplesPlasmid IsolationDNA/RNA Clean Up	Circulating DNA IsolationViral Nucleic acid IsolationgDNA Isolation FFPE DNA/RNA Isolation	Plasmid extractiongel DNA recoverygenomicDNA/RNA extraction viral nucleic acid extractionCirculating DNA extraction	DNA/RNA Clean Up and concentrationDNA/RNA Isolation from low nucleic acid content samplesResearch immuno assays

• The MagPure magnetic-particle technology combines the speed and efficiency of silica-based DNA purification with the convenient handling of magnetic particles. DNA binds to the silica surface of the magnetic particles in the presence of a chaotropic salt. DNA bound to the particles is then efficiently washed, considerably improving the purity of DNA. High-quality DNA is eluted. The automated purification procedure completely removes enzymes, nucleotides, and other contaminants and inhibitors. Purified DNA is suitable for direct use in downstream applications, such as sequencing and microarray analysis.

Magnetic bead nucleic acid purification technology uses nano or micron superparamagnetic material as the matrix, generally black ferric oxide or yellowish brown ferricoxide as the magnetic material. The surface of bead is coated with appropriate functional groups, which can adsorb nucleic acid. Magnetic beads commonly used for nucleic acids, containing carboxyl groups, hydroxyl groups, or silicon groups. Silicon-based magnetic beads are the most common, and its principle of adsorbing nucleic acid is consistent with the classical glass milk purification technology or glass fiber filter membrane purification method. Magpure particle is a kind of polydisperse fast speed silica magnetic beads. The core is ferricoxide, accounting for 50%, and the surface coating is silica, accounting for 50%. The product can be used for plasmid extraction, gel DNA recovery, product purification, genomic DNA and RNA extraction, and viral nucleic acid extraction.

Propargyl-PEG10-t-butyl ester is a crosslinking reagent that can react with azide-bearing compounds or biomolecules via copper catalyzed Click Chemistry reactions. The carboxyl group can be deprotected under acidic conditions. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.

Propargyl-PEG10-t-butyl ester is a crosslinking reagent that can react with azide-bearing compounds or biomolecules via copper catalyzed Click Chemistry reactions. The carboxyl group can be deprotected under acidic conditions. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.