Description

Nucleic acid testing (NAT) is the method of choice for detection and quantification of a wide range of micro organisms. Primerdesign manufactures and supplies high quality quantitative real-time PCR kits for the detection and simultaneous quantification of numerous significant pathogens . A copy number standard curve is provided for quantification and an the internal extraction template (DNA or RNA), controls for the quality of the nucleic acid extraction and eliminates false negative results.

The kit is designed with the broadest possible detection profile to ensure that all clinically relevant strains and subtypes are detected. Target sequences are selected by working with data from key opinion leaders in the field. Multiple sequence alignments and unprecedented real-time PCR expertise in design and validation ensure the best possible kit. Details of the target and priming specificity are included in the individual handbooks above.

Packaged, optimised and ready to use. Expect Better Data.

Primer and probe mix (150 reactions)
Reverse Transcription, target specific primers (RNA genome viruses only)
Copy number standard curve (sufficient for multiple standard curves)
Internal extraction control – Read through VIC channel*
Endogenous control (150 tests)
RNAse/DNAse free water
*alternative fluorophores available on request

DNA Concentrator (Magnetic Beads)

The DNA Concentrator (Magnetic Beads) can be used to efficiently concentrate DNA/RNA from various samples with low concentration without the need of a DNA vacuum concentrator. Solid Phase Reversible Immobilization (SPRI) magnetic beads are well used for DNA and RNA purification and concentration. The beads are paramagnetic particles coated with carboxyl groups that reversibly bind to DNA and RNA.

The concentrator protocol is simple: mix Concentrator Buffer and Concentrator Beads with the sample, wash, and elute pure DNA in a small volume to concentrate the DNA samples. Moreover, the DNA/RNA samples are also purified during the procedure. The beads with our unique technology purify DNA/RNA samples effectively by removing unwanted components such as dNTPs, enzymes, detergents, proteins, and other contaminants.

However, traditional magnetic beads can only bind nucleic acids that are 100 bp or longer. Nucleic acids shorter than 100 bp are not effectively recovered. We have successfully developed the kit that overcomes the hurdle of short nucleic acids recovery. The reagent can be used for concentration of samples from genomic DNA to short DNA/RNA.

Recovery rate

• >90% for DNA size >30 bp
• >80% for DNA size between 20-29 bp

Features

• Effective concentrator of DNA and RNA samples, even small DNA, RNA, oligos
• Binding capacity: 10 ug DNA per prep
• Removal of unwanted components and other impurities

The DNA Concentrator (Magnetic Beads) can be used to efficiently concentrate DNA/RNA from various samples with low concentration without the need of a DNA vacuum concentrator. Solid Phase Reversible Immobilization (SPRI) magnetic beads are well used for DNA and RNA purification and concentration. The beads are paramagnetic particles coated with carboxyl groups that reversibly bind to DNA and RNA.

Introduction

This 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 20mg tissue, 150mg plant, 5 x 106 cell using two columns (gDNA removed column)
Applications	RT-PCR, qRT-PCR, Northern hybridization, second generation sequencing
Purification method	Mini spin column
Purification technology	Silica technology, DNA filtration 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 sample: 1-20 mgPlant tissue: 50-150 mg
Yield	2-100μg
Elution volume	≥50μl
Time per run	≤25 minutes（1-24 samples）
Liquid carrying volume per column	800µl
Binding yield of column	100µg

Principle

The Kit isolates total RNA from up to 10⁷ cells or 20 mg tissue. A short workflow enables RNA isolation with genomic DNA removal in less than 25 mins. 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 a RNA column. RNA binds to the membrane and contaminants are washed away. High-quality RNA is eluted in as little as 30µl water using the Kit.

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 25 minutes by column method
• Safe – no phenol chloroform extraction required
• Sensitive – RNA can be recovered at the level of PG

Kit Contents

Contents	R411102	D411103
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 RW1	50 ml	200 ml
Buffer RW2*	12 ml	2 x 50 ml
RNase Free Water	10 ml	30 ml

Storage and Stability

HiPureKit can be stored dry at room temperature (15-25°C) and are stable for at least18 months under these conditions. During shipment, crystals or precipitationmay form in the Buffer RLC. Dissolve by warming buffer to 37°C.

This 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.