Description

As this is a 2 gene kit, we recommend purchase of 2 of the accompanying RT-qPCR master mix reagent: oasig Lyophilised OneStep RT-qPCR Master Mix 150 reactions.

Norovirus is known to cause acute gastroenteritis. It is a small (27-38 nm), round, nonenveloped RNA virus belonging to the Caliciviridae family and is responsible for over 80% of non-bacterial outbreaks of gastroenteritis in the world. It affects individuals of all ages, with a distinct seasonal link to winter. It has a genome of 7.6 kb that is positive sense and has a single stranded linear confirmation. It encodes a major structural protein (VP1) of about 58 to 60 kDa and a minor capsid protein (VP2). Transmission occurs predominantly through ingestion of contaminated water, food and airborne transmission, as well as contact with contaminated surfaces. The ease with which norovirus is transmitted and the low infectious dose required to establish an infection results in extensive outbreaks in numerous environments, such as hospitals, hotels and schools. There is no antiviral drug available to treat this infection and little is known about its pathogenicity. However, it has been observed that the virus can be taken up by enterocytes where translation of viral nonstructural proteins can occur; it damages and alters intestinal microvilli, leaving them blunt and broadened, thus inhibiting absorption; it causes crypt cell hyperplasia and also leads to apoptosis of enterocyctes. An incubation period of 24-48 hours is usual. Infection is characterized by the acute onset of nausea, vomiting, abdominal cramps, aching limbs, raised temperature and diarrhoea that generally last for about 48 hours. However, more severe and prolonged infection may be observed in children and the elderly. There are five recognized norovirus genogroups, of which three (GI, GII, and GIV) are known to affect humans and, since 2002, variants of the GII.4 genotype have been the most common cause of norovirus outbreaks. There have been 31 different genotypes identified within the genogroups, with a wide degree of genetic variability present even within each genotype.

Genomic DNA Extraction Kit (HMW, Magnetic Beads)

The Genomic DNA Extraction Kit (HMW, Magnetic Beads) provides a reliable and fast process for extracting high molecular weight (HMW) genomic DNA from cells, blood, and tissues using Solid Phase Reversible Immobilization (SPRI) magnetic beads. With our proprietary magnetic beads technology, the kit eliminates the tedious centrifuge steps for columns. The kit provides a reliable and simple approach for high-quality genomic DNA isolation with fast magnetic response time and high binding capacity.

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Cat.# 50014 Genomic DNA Extraction Kit for Cells (HMW, Magnetic Beads)
Cat.# 50015 Genomic DNA Extraction Kit for Blood (HMW, Magnetic Beads)
Cat.# 50016 Genomic DNA Extraction Kit for Tissues (HMW, Magnetic Beads)

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The extracted HMW genomic DNA size ranges are dependent on the beads resuspension: 50-150 kb by tube tapping and 40-100 kb by tube vortexing. Purified DNA is recovered at high yield and high purity without RNA contamination. The typical purity ratios of A260/A280 are around 1.8-2.0, and A260/A230 are around 2.2-2.5. Purified HMW genomic DNA is suitable for applications such as long-read sequencing, linked-read genome assembly, long range PCR, optical mapping, and other general applications.

Features

• High molecular weight DNA: 50 kb to 150 kb
• High purity
• Simple magnetic beads method
  • No centrifuge needed
  • No column needed
  • No vacuum needed

A portion of the extracted genomic DNA samples were loaded on a PFGE gel with a DNA ladder indicated. Sample A: liver tissue; Sample B: intestine tissue; Sample C: whole blood; Sample D: cultured 293T cells.

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Cultured Cell samples

Cultured cells are collected and are resuspended in a buffer and then lysed with a lysis buffer, then mixed with beads to bind genomic DNA. The samples are mixed with a buffer and after washing steps, genomic DNA is eluted in the Elution Buffer. The isolated genomic DNA with the magnetic beads is free of contamination such as RNA, proteins, salts, and other impurities.

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Blood samples

Whole blood is resuspended in the RBC buffer to remove RBC. The remaining leucocytes are lysed with a lysis buffer, then mixed with beads to bind genomic DNA. The samples are mixed with a buffer and after washing steps, genomic DNA is eluted in Elution Buffer. The isolated genomic DNA with the magnetic beads is free of contamination such as RNA, proteins, salts, and other impurities.

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Tissue samples

Tissues are homogenized and lysed, then mixed with beads to bind genomic DNA. The samples are mixed with a buffer and after washing steps, genomic DNA is eluted in Elution Buffer. The isolated genomic DNA with the magnetic beads is free of contamination such as RNA, proteins, salts, and other impurities.

DBCO-PEG2-PFP ester is a PEG active ester consisting of a DBCO group which can react with azides under copper free conditions. The PFP ester is an active ester which can react with amine groups. PFP esters have been are stable compounds and are less susceptible to undergo hydrolysis. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.