Overview

• Sample collection is non-invasive and painless
• Fast and easy high throughput processing using 96-well plates
• Isolate high quality genomic DNA
• Effective removal of PCR inhibitors
• Compatible with preserved saliva samples collected using Norgen’s Saliva DNA Collection and Preservation Devices, as well as fresh saliva samples

This kit features a rapid, high-throughput 96-well format for isolating high yield and high quality DNA from saliva.  The purified saliva DNA is compatible with any number of downstream applications including PCR, qPCR, Southern Blot analysis, sequencing, microarray analysis and more. This kit is also compatible with Norgen’s Saliva DNA Collection and Preservation Devices (Cat. RU49000).  It is fully compatible to both vacuum and centrifugation protocols.

​Background

Human genomic DNA found in saliva can be used in various applications in diagnostics including the detection of biomarkers to diagnose a disease, follow the disease’s progress or monitor the effects of a particular treatment. Isolation of DNA from saliva for diagnostics has become an attractive alternative to isolation from blood or tissue due to the fact that sample collection is non-invasive.

Details

Supporting Data

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Kit Specifications
Binding Capactity Per Well	50 μg
Maximum Loading Volume Per Well	600 μL
Size of DNA Purified	All sizes
Time to Complete 96 Purifications	40 minutes

Storage Conditions and Product Stability
The kit contains ready-to-use Proteinase K which is dissolved in a specially prepared storage buffer. The Proteinase K should be stored at room temperature or 4°C. All other solutions should be kept tightly sealed and stored at room temperature (15 – 25°C). This kit is stable for 1 year after the date of shipment.

Component	Cat. RU35200 (192 preps)
Lysis Buffer F	100 mL
Proteinase K in Storage Buffer	4 mL
Binding Buffer B	40 mL
Wash Solution A	2 x 38 mL
Elution Buffer B	30 mL
96-Well Plate	2
96-Well Collection Plate	2
Adhesive Tape	4
96-Well Elution Plate	2
Product Insert	1

Introduction

With the development of molecular biology, stool, a new non-invasive sample, has been widely used in the research of animal molecular genetics, population ecology, behavioral ecology and some intestinal disease diagnosis. Stool samples includes gut microbial DNA, food residue sample DNA, and alimentary tract exfoliated cell DNA.

The primary problem encountered when using stool sample for molecular biology research is the low content of exfoliated cells in the digestive tract and a certain degree of degradation of genetic material in stool. Another issue in molecular scatology research based on PCR is the presence of a large number of inhibitors in stool that can affect Taq enzyme activity, leading to downstream detection inactivation. These inhibitors include polysaccharides, plant polysaccharides, bile acids, bile salts, bile pigments, digestive juices, mucus, etc. Therefore, selecting appropriate extraction methods to obtain high-quality DNA is the key to successful downstream detection of stool DNA.

At present, the pretreatment methods used in the laboratory, such as phenol/chloroform extraction, cetyltrimethyl bromide (CTAB) lysis, and guanidine isothiocyanate lysis, lack universality in different species, and the success rate of extracting DNA for PCR amplification is also very low. The HiPure Stool DNA Kit provided by Magen Company has opened up a new approach for DNA extraction from stool samples with good universality, high cost-effectiveness, high yield and purification. The reagent kit adopts a unique solution system and inhibitory factor adsorbent, which can efficiently remove various impurities in stool samples. The purified DNA can be directly used for PCR, quantitative PCR and other applications.

This product allows rapid and reliable isolation of high-quality genomic DNA from various stool samples. Up to 100 mg soil samples can be processed in 60 minute. The system combines the reversible nucleic acid binding properties of HiPure matrix with the speed and versatility of spin column technology to eliminate PCR inhibiting compounds such as humic acid from soil samples. Purified DNA is suitable for PCR, restriction digestion, and next-generation sequencing. There are no organic extractions thus reducing plastic waste and hands-on time to allow multiple samples to be processed in parallel.

Details

Specifications

Features	Specifications
Main Functions	Isolation total DNA from 50-100mg stool samples
Applications	PCR, Southern Blot, enzyme digestion and NGS, etc.
Purification method	Mini spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	Stool
Sample amount	50-100mg
Yield	3-15μg
Elution volume	≥30μl
Time per run	≤60 minutes
Liquid carrying volume per column	750μl
Binding yield of column	100μg

Principle

Stool sample is homogenized and then treated in a specially formulated buffer containing detergent to lyse bacteria, yeast, and fungal samples. Humic acid, proteins, polysaccharides, and other contaminants are removed using our proprietary Absorber Solution. Binding conditions are then adjusted and the sample is applied to a DNA Mini Column. Two rapid wash steps remove trace contaminants and pure DNA is eluted in low ionic strength buffer. Purified DNA can be directly used in downstream applications without the need for further purification.

Advantages

• High purity – unique adsorbent can completely remove inhibitory factors
• High concentration – maximum extraction of total DNA from stool samples
• High recovery – DNA can be recovered at the level of PG
• Good repeatability – silica technology can obtain ideal results every time

Kit Contents

Contents	IVD3141
Purification Times	50 Preps
HiPure DNA Mini Columns II	50
2ml Collection Tubes	50
2ml Bead Tubes	50
Proteinase K	24 mg
Protease Dissolve Buffer	1.8 ml
Buffer SPL	40 ml
Buffer PCI	40 ml
Buffer AL	20 ml
Buffer GW1	22 ml
Buffer GW2	20 ml
Buffer AE	15 ml

Storage and Stability

Proteinase K and Buffer PCI 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 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.

With the development of molecular biology, stool, a new non-invasive sample, has been widely used in the research of animal molecular genetics, population ecology, behavioral ecology and some intestinal disease diagnosis. Stool samples includes gut microbial DNA, food residue sample DNA, and alimentary tract exfoliated cell DNA.

Overview

• Detection kits for the BKV/JCV
• Available in TaqMan format for analysis

The BK virus is a member of the polyomavirus family. It has been suggested that this virus may be transmitted through respiratory fluids or urine, since infected individuals periodically excrete virus in the urine. BK viral infections are typically asymptomatic in healthy individuals, however very mild symptoms may appear including mild respiratory infections and fever. Infections with BK virus in immunocompromised or immunosupressed patients are much more severe and may involve renal dysfunction. In fact, in kidney transplant patients the immunosupressive drugs required for the transplant may allow the virus to replicate within the graft, resulting in a disease called BK virus nephropathy (BKVN). The JC virus is a type of human polyomavirus and is very common in the general population, infecting 70 to 90% of humans. Most people acquire JCV in childhood or adolescence. Typically the infection is subclinical and no of consequence in individuals with healthy immune systems. The initial site of infection may be the tonsils or the gastrointestinal tract, and the virus then remains latent in the gastrointestinal tract. JCV can also infect the tubular epithelial cells in the kidneys, where it continues to reproduce, shedding virus particles in the urine. Also, JCV can cross the blood-brain barrier into the central nervous system. JCV is known to cause the usually fatal progressive multifocal leukoencephalopathy (PML) by destroying oligodendrocytes in the brain in immunodeficient or immunosuppressed individuals. The JC and BK viruses are very similar, with their genomes sharing 75% homology. It is however important to differentiate between the viruses due to the differences in pathology and especially the invariably fatal outcome of PML which is only caused by the JC virus.

BKV/JCV TaqMan PCR Kit, 100 reactions

• Ready to use format, including Master Mix for the target and PCR control to monitor for PCR inhibition and validate the quality
• Specific Primer and Probe mix for the pathogen/virus/viroid of interest
• Primer and Probe mix
• Positive and negative control to confirm the integrity of the kit reagents

BKV/JCV TaqMan PCR Probe/Primer Set and Controls, 100 reactions

• Specific Primer/Probe mix and Positive Control for the pathogen/virus/viroid of interest
• Nuclease-free water
• Can be used together with Norgen’s PCR Master Mix (#28007) or customer supplied master mix

For research use only and NOT intended for in vitro diagnostics.

Details

Supporting Data

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Storage Conditions and Product Stability
All kit components can be stored for 2 years after the date of production without showing any reduction in performance.

All kit components should be stored at -20°C upon arrival. Repeated thawing and freezing (> 2 x) of the Master Mix and Positive Control should be avoided, as this may affect the performance of the assay. If the reagents are to be used only intermittently, they should be frozen in aliquots.

Component	Cat. TM39350 (100 preps)	Cat. TM39310 (100 preps)
MDx TaqMan 2X PCR Master Mix	2 x 700 μL	–
BKV/JCV Primer & Probe Mix	280 μL	280 μL
BKV/JCV Positive Control	150 μL	150 μL
Nuclease-Free Water (Negative Control)	1.25 mL	1.25 mL
Product Insert	1	1