Description

Nucleic acid testing (NAT) is the method of choice for detection and quantification of a wide range of micro organisms. Primerdesign manufactures and supplies high quality quantitative real-time PCR kits for the detection and simultaneous quantification of numerous significant pathogens . A copy number standard curve is provided for quantification and an the internal extraction template (DNA or RNA), controls for the quality of the nucleic acid extraction and eliminates false negative results.

The kit is designed with the broadest possible detection profile to ensure that all clinically relevant strains and subtypes are detected. Target sequences are selected by working with data from key opinion leaders in the field. Multiple sequence alignments and unprecedented real-time PCR expertise in design and validation ensure the best possible kit.

Details of the target and priming specificity are included in the individual handbooks above.

Packaged, optimised and ready to use. Expect Better Data.

Exceptional value for money
Rapid detection of all clinically relevant subtypes
Positive copy number standard curve for quantification
Highly specific detection profile
High priming efficiency
Broad dynamic detection range (>6 logs)
Sensitive to < 100 copies of target Accurate controls to confirm findings

Introduction

This product provides an easy-to-use workflow for selective isolation of bacterial DNA from samples that are intrinsically rich in host DNA, such as body fluids or swabs. The method is specific for the identification of intact bacteria so it prevents false results due to nucleic acids from dead bacteria. The Kit allows isolation of enriched bacterial DNA suitable for a variety of applications, including qPCR and whole metagenome or 16S rRNA gene sequencing.

Details

Specifications

Features	Specifications
Main Functions	Isolation gDNA from biological sample and remove host DNA
Applications	PCR, southern blot and enzyme digestion, etc.
Purification method	Mini spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	Culture medium, swab, parasitic blood, tissue, sputum, etc.
Sample amount	Blood sample, homogenate, plasma, brain effusion, swab immersion solution, centrifuged concentrated liquid, etc.:0.5-1ml
Elution volume	≥50μl
Time per run	≤60 minutes
Liquid carrying volume per column	800μl
Binding yield of column	100μg

Principle

This product is based on silica Column purification. This product efficiently depletes human and animal host DNA and yields enriched bacterial DNA. An optimized combination of mechanical and chemical lysis allows efficient disruption of bacterial cells. Target DNA 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 (10mm Tris, pH9.0, 0.5mm EDTA).

Advantages

• High concentration – the host DNA is digested by dnase, without contamination of host DNA
• High recovery – DNA can be recovered at the level of PG
• Good repeatability – silica technology can obtain ideal results every time
• Wide applicability – it can be used in blood, tissue, intestinal contents and other samples

Kit Contents

Contents	D314802	D314803
Purification Times	50 Preps	250 Preps
Hipure DNA Mini Columns I	50	2 x 125
2ml Collection Tubes	50	2 x 125
2ml beads Tubes	50	250
Buffer DRB	15 ml	60 ml
Buffer ES	6 ml	30 ml
Reagent DX	0.5 ml	1 ml
Buffer DL	30 ml	120 ml
Buffer GW1	22 ml	110 ml
Buffer GW2	12 ml	50 ml
DNase I (Powder)	6 mg	30 mg
Proteinase K	24 mg	120 mg
Protease Dissolve Buffer	5 ml	15 ml
Buffer AE	15 ml	60 ml

Storage and Stability

DNase I 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 their performance. The remaining kit components can be stored dry at room temperature (15–25°C) and are stable for at least 18 months under these 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 product provides an easy-to-use workflow for selective isolation of bacterial DNA from samples that are intrinsically rich in host DNA, such as body fluids or swabs. The method is specific for the identification of intact bacteria so it prevents false results due to nucleic acids from dead bacteria. The Kit allows isolation of enriched bacterial DNA suitable for a variety of applications, including qPCR and whole metagenome or 16S rRNA gene sequencing.

Overview

• Purification and enrichment of intact exosomes from plasma, serum, urine, cell culture media and saliva in less than 30 minutes.
• Versatile sample input ranging from 50 µL to 10 mL
  • Plasma/Serum Exosome Purification Mini Kit (50 µL – 1 mL Plasma/Serum)
  • Plasma/Serum Exosome Purification Midi Kit (1 mL – 4 mL Plasma/Serum)
  • Plasma/Serum Exosome Purification Maxi Kit (4 mL – 10 mL Plasma/Serum)
• Exosome purification is based on Norgen’s proprietary resin separating matrix through exosomes’ surface proteins.
• No precipitation reagents, overnight incubation, protease or coagulant treatments required
• No time-consuming ultracentrifugation, filtration or special syringes required
• Purify intact exosomes with a size ranging from 40-200 nm depending on sample input type
• Purified exosomes are compatible with functional studies.
• Purified exosomes are free from any RNA-binding proteins
• Purified exosomes are compatible with NanoSight® or Electron Microscopy for assessing the approximate exosome size range and concentration.
• Exosomal RNA can be extracted from the purified exosomes using Norgen’s Exosomal RNA Purification technology or any other RNA extraction method.

The Plasma/Serum Exosome Purification Kits provide a fast, reliable and convenient method to purify and enrich for intact exosomes from different plasma/serum sample volumes ranging from 50 µL to 10 mL. These kits also allow for the purification of intact extracellular vesicles (EVs) from different plasma/serum sample volumes, and these EVs are ready for any downstream application. The purification is based on Norgen’s proprietary resin.

These kits provide a clear advantage over other available methods since they do not require any special instrumentation, ultracentrifugation, precipitation reagents or any protease treatments. More importantly, the purified exosomes will not be contaminated with any other RNA-binding proteins that may contaminate your exosomal RNA, which is essential if studying exosomal transcripts.

NanoSight® Analysis

Exosomes enriched with Norgen’s Plasma/Serum Exosome Purification Kits can be analyzed using NanoSight® for assessing the approximate exosome size range and concentration

Exosomal RNA Analysis

To purify exosomes and isolate exosomal RNA, choose the Plasma/serum Exosome Purification and RNA isolation kits. The protocol is divided into 2 parts and an aliquot of purified exosomes can be taken for applications like NTA/TEM etc. before processing them for RNA isolation. Or you can use the Exosomal RNA Isolation Kit if you’ve already purified exosomes using a Norgen kit or another method. . Exosomal RNA isolation is based on Norgen’s proprietary resin without the need for phenol extractions or carrier RNA. This RNA is ideal for gene expression analysis using RT-qPCR, microarray, or NGS and for biomarker discovery.

Details

Supporting Data

Figure 1 / 4

Previous

Next

Click for expanded view

Kit Specifications
Plasma/Serum Input (Cat. 57400)	50 μL – 1 mL
Plasma/Serum Input (Cat. 57500)	1 mL – 4 mL
Plasma/Serum Input (Cat. 57600)	4 mL – 10 mL
Size of Exosomes Purified	40 nm – 150 nm
Elution Volume	Variable depending on the plasma/serum input volume
Time to Complete 10 Purifications	15 – 30 minutes

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

Important Note
This kit is suitable for the purification of exosomes from fresh or frozen serum or plasma prepared from blood collected on either EDTA or Citrate. Plasma samples prepared from blood collected on heparin should not be used as heparin can significantly interfere with many downstream applications such as RT-PCR.

Component	Cat. 57400 (50 preps)	Cat. 57500 (25 preps)	Cat. 57600 (15 preps)
Slurry E	12.5 mL	12.5 mL	12.5 mL
ExoC Buffer	8 mL	8 mL	2 x 8 mL
ExoR Buffer	12 mL	12 mL	12 mL
Mini Filter Spin Columns inserted into 2 mL tubes	50	25	15
Product Insert	1	1	1