Introduction

The HiPure Total RNA Kit provides fast purification of high-quality RNA from cells, tissues, and yeast 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. RNA purified using the HiPure Total RNA Purification System 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 (not include miRNA) from animal tissues, cells and simple plant tissues using one column
Applications	RT-PCR, qRT-PCR, Northern hybridization, second generation sequencing
Purification method	Mini spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	Animal soft tissue, cultured cells, lymphocytes, simple plant tissue
Sample amount	Cells: ≤1 x 107 Animal tissue: 1-20 mgPlant leaves: 50-150 mg Yeast cells: 5 x 106
Yield	2-100μg
Elution volume	≥50μl
Time per run	≤15 minutes（1-24 samples）
Liquid carrying volume per column	800µl
Binding yield of column	100µg

Principle

HiPure RNA technology simplifies total RNA isolation. Samples are first lysed and then homogenized. Ethanol is added to the lysate to provide ideal binding conditions. The lysate is then loaded onto the HiPure silica membrane and RNA binds to the silica membrane, and all contaminants are efficiently washed away. For certain RNA applications that are sensitive to very small amounts of DNA, the residual amounts of DNA remaining can be removed using a convenient on-column DNase treatment. Pure, concentrated RNA is eluted in water.

Advantages

• Efficient removal of DNA – unique genomic DNA removal column without DNase treatment
• High quality – high purity total RNA can be directly used in various sensitive downstream applications
• Fast – several samples can be extracted in 20 minutes by column method
• Safe – no phenol chloroform extraction required
• Sensitive – RNA can be recovered at the level of PG

Kit Contents

Contents	R401102	R401103
Purification Times	50 Preps	250 Preps
HiPure RNA Mini Columns	50	250
2ml Collection Tubes	50	250
RTL Lysis Buffer	50 ml	200 ml
RNA Binding Buffer	15 ml	75 ml
Buffer RW1	50 ml	200 ml
Buffer RW2*	20 ml	2 x 50 ml
RNase Free Water	10 ml	30 ml

Storage and Stability

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

The HiPure Total RNA Kit provides fast purification of high-quality RNA from cells, tissues, and yeast 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. RNA purified using the HiPure Total RNA Purification System can be used for applications such as RT-PCR, Northern blotting, poly A+ RNA (mRNA) purification, nuclease protection, and in vitro translation.

Description 

The ExcelTaq™ 2X Q-PCR Master Mix (SYBR, no ROX) is a ready-to-use reagent with all the essential components for quantitative real-time PCR (qPCR) except primers and template. The master mix features high sensitivity and signal intensity as well as low background and better compatibility with cDNA templates derived directly from reverse transcription reaction mixture. The ExcelTaq™ 2X Q-PCR Master Mix (SYBR, no ROX) contains hot-start Taq polymerase in an optimized buffer with dsDNA specific SYBR green fluorescent dye. This master mix allows for sensitive, precise amplification, real-time tracking of the amplification process, and simultaneous quantification for targeted DNA molecules. With inert smart blue contrast dye, the ExcelTaq™ 2X Q-PCR Master Mix (SYBR, no ROX) is ready-to-use and greatly reduces pipetting errors, while largely improving the reproducibility of the process. The ExcelTaq™ 2X Q-PCR Master Mix is also compatible with a ROX reference dye if recommended by the manufacturer of the qPCR system. 

Features

• High sensitivity and signal intensity
• Smart blue contrast dye as a visual aid for reaction setup
• Better compatibility for reverse transcription
• Low background 

Storage 

Protected from light.
Aliquot to avoid multiple freeze-thaw cycles.
-20°C for 12 months

The ExcelTaq™ 2X Q-PCR Master Mix (SYBR, no ROX) is a ready-to-use reagent with all the essential components for quantitative real-time PCR (qPCR) except primers and template. The master mix features high sensitivity and signal intensity as well as low background and better compatibility with cDNA templates derived directly from reverse transcription reaction mixture. The ExcelTaq™ 2X Q-PCR Master Mix (SYBR, no ROX) contains hot-start Taq polymerase in an optimized buffer with dsDNA specific SYBR green fluorescent dye. This master mix allows for sensitive, precise amplification, real-time tracking of the amplification process, and simultaneous quantification for targeted DNA molecules. With inert smart blue contrast dye, the ExcelTaq™ 2X Q-PCR Master Mix (SYBR, no ROX) is ready-to-use and greatly reduces pipetting errors, while largely improving the reproducibility of the process. The ExcelTaq™ 2X Q-PCR Master Mix is also compatible with a ROX reference dye if recommended by the manufacturer of the qPCR system.