Overview

• Ensure optimal results for sensitive applications like NGS
• Up to a tenfold increase in microRNA mapping during sequencing runs to reduce costs
• Isolates all sizes of exosomal and extracellular vesicle RNA, including microRNA.
• Bind and elute all RNA irrespective of size or GC content, without bias.
• No phenol extractions.
• No Proteinase K treatment.
• No carrier RNA.
• Concentrate isolated RNA into a flexible elution volume ranging from 50 μL to 100 μL.
• Purify high-quality RNA in 15-20 minutes.
• Purified RNA is suitable for a variety of downstream applications, including Small RNA Sequencing.
• Purification is based on EXTRAClean spin column chromatography that uses Norgen’s proprietary separation matrix.

The kit is designed to isolate all sizes of RNA, including microRNA. The kit provides a clear advantage over other available kits in that it does not require any special instrumentation, protein precipitation reagents, extension tubes, phenol/chloroform or protease treatments. Moreover, the kit allows the user to elute into a flexible elution volume ranging from 50 μL to 100 μL. The purified RNA is of the highest integrity, and can be used in a number of downstream applications including real time PCR, reverse transcription PCR, Northern blotting, RNase protection and primer extension, and expression array assays.

Details

Supporting Data

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Sample Type	Exosomes purified using Norgen’s Purification Kits
Size of RNA Purified	All sizes, including miRNA and small RNA (< 200 nt)
Elution Volume	50-100 μL
Time to Complete 10 Purifications	35-40 minutes
Average Yields	Variable depending on specimen

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. It is recommended to warm Lysis Buffer A for 20 minutes at 60°C if any salt precipitation is observed.

Introduction

The Kit provides fast purification of high-quality DNA, RNA and Prote from cells, tissues, and yeast using silica-membrane spin columns with a binding capacity of 100ug RNA. There is no need for phenol/chloroform extractions and time-consuming steps such as CsCl gradient ultracentrifugation, or precipitation with isopropanol or LiCl are eliminated. RNA purified using the HiPure Total RNA Purification System 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	Co-isolation DNA and RNA /protein from a single sample (cells, soft tissue, plant sample)
Applications	SDS-PAGE electrophoresis and western blot, etc
Purification method	Mini spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	Culture cells, animal tissues, plant fungi, yeast, bacteria and other samples
Sample amount	Cultured cells: < 10^7Animal tissue: ≤ 20 mgPlant samples: ≤ 150 mgYeast cells: 2 x 10^6 – 5 x 10^7

Principle

The Kit isolates total RNA from up to 10 ⁷ cells or 30 mg tissue. A short workflow enables RNAisolation with genomic DNA removal in less than 25 min. Samples are first lysed and homogenized. The lysate is passed through a DNA Mini column, ethanol is added to the flow-through, and the sample is applied to an RNA column. RNA binds to the membrane and contaminants are washed away. High-quality RNA is eluted in as little as 30 µl water using the Kit.

Advantages

• High quality – high purity total RNA / DNA can be directly used in a variety of downstream applications
• Fast – column method can complete the extraction of several samples in 30 minutes
• Safe – no phenol chloroform extraction required
• Simultaneous extraction- simultaneously isolate DNA and RNA from one sample

Kit Contents

Contents	R521102	R521103
Purification Times	50 Preps	250 Preps
HiPure DNA Mini Columns	50	250
HiPure RNA Mini Columns	50	250
2ml Collection Tubes	100	2 x 250
Buffer RLC	50 ml	200 ml
Buffer GW1*	22 ml	66 ml
Buffer RW1	50 ml	200 ml
Buffer RW2*	50 ml	3 x 50  ml
RNase Free Water	10 ml	30 ml
Elution Buffer	10 ml	30 ml
Buffer ALO(5%SDS)	10 ml	30 ml

Storage and Stability

HiPure Kit can be stored dry at room temperature (15–25°C) and are stable for at least 18 months under these conditions. During shipment, crystals or precipitation may form in the Buffer RLC. Dissolve by warming buffer to 37°C.

The Kit provides fast purification of high-quality DNA, RNA and Prote from cells, tissues, and yeast using silica-membrane spin columns with a binding capacity of 100ug RNA. There is no need for phenol/chloroform extractions and time-consuming steps such as CsCl gradient ultracentrifugation, or precipitation with isopropanol or LiCl are eliminated. RNA purified using the HiPure Total RNA Purification System is ready for applications such as RT-PCR, Northern blotting, poly A+ RNA (mRNA) purification, nuclease protection, and in vitro translation.

Overview

• Detection kits for Cryptococcus neoformans
• Available in TaqMan format for analysis

Cryptococcus neoformans is an encapsulated yeast. Infection with C. neoformans is known as cryptococcosis and is the cause of the most common life-threatening meningitis in patients with weakened immune systems, particularly in advanced HIV/AIDS. C. neoformans is commonly found in soil throughout the world. Human infection of C. neoformans occurs via inhalation of aerosolized spores. From the lungs, C. neoformans is spread hematogenously to the Central Nervous System (CNS), resulting in meningoencephalitis. Although the availability of antiretroviral therapy in the developed world has reduced the incidents of cryptococcosis, it is still a major problem in developing countries and is one of the leading causes of death in patients with HIV/AIDS. In fact, one of the major challenges in treating cryptococcosis is that many patients with cryptococcal CNS disease are asymptomatic in terms of cryptococcal pneumonia, making it difficult for early detection.

Cryptococcus neoformans 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

Cryptococcus neoformans 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

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. TM42750 (100 preps)	Cat. TM42710 (100 preps)
MDx TaqMan 2X PCR Master Mix	2 x 700 μL	–
Cryptococcus neoformans Primer & Probe Mix	280 μL	280 μL
Cryptococcus neoformans Positive Control	150 μL	150 μL
Nuclease-Free Water (Negative Control)	1.25 mL	1.25 mL
Product Insert	1	1