Introduction

The Kit provides fast purification of high-quality RNA from whole blood, cells and tissues using silica-membrane spin columns with a binding capacity of 100μg RNA. There is no need for phenol / chloroform extractions and time-consuming steps such as CsCl gradient ultracentrifugation or isopropanol precipitation. The purified RNA can be used 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 from 1-1.5ml whole blood
Applications	qPCR / RT-PCR, liquid or solid-phasechip analysis, hybridization and SNP detection
Purification method	Mini spin column
Purification technology	Silica technology, acidphenol / guanidine extraction technology (MagZol pretreatment technology)
Process method	Manual (centrifugation or vacuum)
Sample type	Fresh or frozen blood, mammalian blood
Sample amount	≤1.5 ml whole blood
Yield	2-100μg
Elution volume	≥20μl
Liquid carrying volume per column	800µl
Binding yield of column	100μg

Principle

The Kit simplifies isolation of RNA from blood with a fast spin-column procedure. Red blood cells are selectively lysed and white cells collected by centrifugation. White cells are then lysed using highly denaturing conditions which immediately inactivate RNases. After homogenization using the DNA spin column, the sample is applied to the RNA column. Total RNA binds to the membrane and contaminants are washed away, leaving pure RNA to be eluted in 30–100µl RNase-free water (provided with the kit) for direct use in any downstream application.

Advantages

• High quality – high purity total RNA can be directly used in various sensitive downstream applications
• Fast – isolation of several samples can be completed in 20 minutes by using column purification method
• Sensitive – RNA can be purified at the level of PG

Kit Contents

Contents	R416102	R416103
Purification Times	50 Preps	250 Preps
HiPure DNA Mini Columns	50	250
HiPure RNA Mini Columns I	50	250
2ml Collection Tubes	100	500
10 x Buffer RBC	50 ml	3 x 100 ml
RTL Lysis Buffer	50 ml	250 ml
Buffer RW1	50 ml	250 ml
Buffer RW2*	20 ml	2 x 50 ml
RNase Free Water	10 ml	30 ml

Storage and Stability

HiPure Blood RNA Mini Kit components can be stored dry at room temperature (15-25°C) and are stable for at least 18 months under these conditions. Make sure that all buffers are at room temperature when used. During shipment, crystals or precipitation may form in the RTL Lysis Buffer. Dissolve by warming buffer to 37°C.

The Kit provides fast purification of high-quality RNA from whole blood, cells and tissues using silica-membrane spin columns with a binding capacity of 100μg RNA. There is no need for phenol / chloroform extractions and time-consuming steps such as CsCl gradient ultracentrifugation or isopropanol precipitation. The purified RNA can be used for applications such as RT-PCR, Northern blotting, poly A+ RNA (mRNA) purification, nuclease protection and in vitro translation.

Overview

• Quantified standard to be used as a positive control or PCR quantification standard
• Vigorously quantified using multiple methods

Mastitis is the single most costly disease of dairy cattle resulting in the reduction of milk yield and quality. The inflammation of the utter is mainly caused by bacteria, and Staphylococcus aureus (S. aureus) is often considered the most common cause of contagious mastitis in dairy herds. S. aureus infection is estimated to be present in up to 90% of dairy farms and is responsible for 35% of the economic loss in the dairy industry. S. aureus is a facultatively anaerobic, Gram positive bacterium. The majority of S. aureus strains are catalase-positive and coagulase-positive, which forms the basis of traditional identification methodology.

Staphylococcus aureus Quantified Bacterial DNA Standard is prepared from cultured bacteria using Norgen’s sample preparation technology. The purified DNA is quantified vigorously using multiple methods including spectrophotometry, gel densitometry and real-time PCR. It is intended to be used as a positive control or PCR quantification standard for Staphylococcus aureus.

Details

Supporting Data

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Volume Provided	250 µL
DNA Quantity	2 x 104 copies per µL

Storage Conditions

Upon receipt, store Norgen’s Staphylococcus aureus Quantified Bacterial DNA Standard at -20^oC or lower. Avoid multiple freeze-thaw cycles. If needed, prepare smaller working aliquots and store at -20^oC or lower.