Overview

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

The BK virus is a member of the polyomavirus family. BK viral infections are typically asymptomatic in healthy individuals, however very mild symptoms may appear including mild respiratory infections and fever. Once an individual has been infected the virus disseminates to the kidneys and the urinary tract where it remains for the lifetime of the individual. 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). It is thought that 1-10% of renal transplant patients progress to BK virus nephropathy (BKVN) and up to 80% of these patients are reported to have lost their grafts. The onset of nephritis can occur as early as several days post-transplant to as late as 5 years. The mode of transmission of the virus is not clear, however it has been suggested that BKV may be transmitted through respiratory fluids or urine, since infected individuals periodically excrete virus in the urine. This virus can be diagnosed by BKV blood & urine testing, in addition to carrying out a biopsy in the kidneys. PCR techniques are now widely used to identify the virus.

BKV 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 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. TM36550 (100 preps)	Cat. TM36510 (100 preps)
MDx TaqMan 2X PCR Master Mix	2 x 700 μL	–
BKV Primer & Probe Mix	280 μL	280 μL
BKV Positive Control	150 μL	150 μL
Nuclease-Free Water (Negative Control)	1.25 mL	1.25 mL
Product Insert	1	1

Introduction

The kit offers the unique feature to isolate total RNA including small RNA and DNA from serum and plasma without the need to resort to the cumbersome phenol/chloroform extraction or a time consuming proteinase digest. RNA purified using the kit is ready for applications such as RT-PCR, Northern blotting, poly A+ RNA (mRNA) purification, nuclease protection, and in vitro translation.

Details

Specifications

Features	Specifications
Main Functions	Isolation total RNA and miRNA from 0.2-0.6ml serum and plasma
Applications
Purification method	Mini spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type
Sample amount
Yield
Elution volume
Time per run
Liquid carrying volume per column
Binding yield of column

Principle

HiPure RNA technology simplifies total RNA isolation. The sample material is denatured in Lysis Buffer. The protein is then precipitated by Protein Precipitation Solution and pelleted by centrifugation. After the removal of protein the binding conditions for nucleic acids are adjusted by adding isopropanol. Total nucleic acids are bound to the column. Optionally, DNA can be removed by an on-column DNase digest. The remaining nucleic acids are washed and eluted with minimal amounts of RNase-Free water.

Kit Contents

Contents	R431402	R431403
Purification Times	50 Preps	250 Preps
HiPure RNA Mini Columns	100	2 x 250
2ml Collection Tubes	100	2 x 250
Buffer CFL	6 ml	30 ml
Buffer CFP	1.8 ml	10 ml
Buffer RWC	20 ml	80 ml
Buffer RW2*	20 ml	2 x 50 ml
RNase Free Water	10 ml	60 ml

Storage and Stability

The kit components can be stored at room temperature (15-25°C) and are stable for at least 18 months under these conditions.

The kit offers the unique feature to isolate total RNA including small RNA and DNA from serum and plasma without the need to resort to the cumbersome phenol/chloroform extraction or a time consuming proteinase digest. RNA purified using the kit is ready for applications such as RT-PCR, Northern blotting, poly A+ RNA (mRNA) purification, nuclease protection, and in vitro translation.

Introduction

HiPure Gel Pure DNA Kit uses proprietary chemistry and HiPure technology to recover DNA fragments between 60bp-10kbp with yields exceeding 80%. DNA is suitable for ligations, PCR, sequencing, restriction digestion, or various labeling reactions. In addition, this kit can be also used to recover DNA directly from enzymatic reactions such as PCR and enzyme digestion reactions.

Details

Specifications

Features	Specifications
Main Functions	Recover DNA fragments >100bp from agarose gel(<0.5g), purification of DNA from PCR, enzymatic reaction solution or crude gDNA
Applications	PCR, NGS, labeling, ligation and enzyme digestion, etc.
Purification method	Mini spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	Agarose gel, PCR products, enzyme products
Sample amount	Agarose gel: ≤500mg
Recovery	≥80%
Elution volume	≥15μl
Time per run	≤20 minutes（1-24 samples）
Liquid carrying volume per column	800µl
Binding yield of column	35µg

Principle

The HiPure system uses a simple bind-wash-elute procedure. Gel slices are dissolved in a buffer containing a pH indicator, allowing easy determination of the optimal pH for DNA binding, and the mixture is applied to the column. Nucleic acids adsorb to the silica-gel membrane in the high-salt conditions provided by the buffer. Impurities are washed away and pure DNA is eluted with a small volume of low-salt buffer provided or water, ready to use in subsequent applications.

Advantages

• High recovery efficiency – ≥80% DNA recovery
• General – recover DNA from gel or enzyme-driven reaction solutions such as PCR
• Fast – isolation can be completed in 10-15 minutes by column gel method
• Great cost-effectiveness performance

Kit Contents

Contents	D211102	D211103
Purification Times	100 Preps	250 Preps
Buffer GDP	120 ml	250 ml
Buffer DW2	50 ml	2 x 50 ml
Elution Buffer	20 ml	30 ml
HiPure DNA Mini Columns II	100	250
2 ml Collection Tubes	100	250

Storage and Stability

The Kit should be stored dry at room temperature (15-25°C) and are stable for at least 18 months under these conditions. If any precipitates form in the buffers, warm at 37℃ to dissolve.

Experiment Data

HiPure Gel Pure DNA Kit uses proprietary chemistry and HiPure technology to recover DNA fragments between 60bp-10kbp with yields exceeding 80%. DNA is suitable for ligations, PCR, sequencing, restriction digestion, or various labeling reactions. In addition, this kit can be also used to recover DNA directly from enzymatic reactions such as PCR and enzyme digestion reactions.