NGS Library Quantification Standards With PCR Primers (Ion Torrent Platform)
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The NGS Library Quantification Standards with PCR Primers (for Ion Torrent platform) were developed for quantifying the library concentration for ion torrent sequencing platform. Quantification of the library of the fully ligated libraries is important for the quality of the sequencing outcome. Optimal library concentration can increase sequencing yield. Poor library concentration results in bad emPCR, which can lead to low sequencing capacity.
Detail
The NGS Library Quantification Standards with PCR Primers (for Ion Torrent platform) were developed for quantifying the library concentration for ion torrent sequencing platform. Quantification of the library of the fully ligated libraries is important for the quality of the sequencing outcome. Optimal library concentration can increase sequencing yield. Poor library concentration results in bad emPCR, which can lead to low sequencing capacity.
QPCR is the best method for library quantification. Our reagent only amplifies library molecules that will be used for subsequent emPCR, and is optimized for amplification of various samples. Our reagent is compatible with commercial SYBR Green based QPCR reagents. Quantification of library concentration is achieved by comparison with a standard curve generated from DNA Standards.
The kit comprises DNA Standards (six 10-fold dilutions) and a primer mix.
NGS Library Quantification Standards with PCR Primers (Ion Torrent platform): real time quantitative PCR curve of the standards.
Other Products
EXTRAClean Plasma/Serum RNA Purification Kit
Product Info
Document
Product Info
Overview
Versatile plasma/serum input ranges
No phenol extractions
Fast and easy processing of samples for the purification and the isolation of RNA.
These kits provide a fast, reliable and convenient method to purify and concentrate high quality, high purity and inhibitor-free cell-free circulating and exosomal RNA using a convenient spin column method. These kits can purify RNA from fresh or frozen serum or plasma samples prepared from blood collected on either EDTA or Citrate. Plasma samples prepared from blood collected on heparin should not be used, as heparin can significantly interfere with many downstream applications such as RT-PCR. The purified plasma/serum RNA is fully compatible with all downstream applications including PCR, qPCR, methylation-sensitive reverse transcription qPCR, reverse transcription PCR, Northern blotting, RNase protection and primer extension, expression array assays, and NGS. The EXTRAClean columns undergo stringent processing and rigorous quality control measures to minimize contamination traces, ensuring optimal results for sensitive applications such as NGS.
Background
Cell-free circulating RNA, including exosomal RNA in plasma or serum, has the potential to provide biomarkers for certain cancers and disease states, and includes tumor-specific extracellular RNA in the blood. Exosomes are 40 – 100 nm membrane vesicles, which are secreted by most cell types. Exosomes can be found in saliva, blood, urine, amniotic fluid and malignant ascitic fluids, among other biological fluids. Evidence has been accumulating recently that these vesicles act as cellular messengers, conveying information to distant cells and tissues within the body. The exosomes contain cell-specific proteins, lipids and RNAs, which are transported to other cells, where they can alter function and/or physiology. These exosomes may play a functional role in mediating adaptive immune responses to infectious agents and tumours, tissue repair, neural communication and transfer of pathogenic proteins. Recent work has demonstrated the presence of distinct subsets of microRNAs within exosomes which depend upon the tumour cell type from which they are secreted. For this reason, exosomal RNAs may serve as biomarkers for various diseases including cancer. As the RNA molecules encapsulated within exosomes are protected from degradation by RNAses, they can be efficiently recovered from biological fluids, such as plasma or serum.
EXTRAClean Plasma/Serum RNA Purification Mini Kit
This kit can purify RNA from fresh or frozen serum or plasma samples prepared from blood collected on either EDTA or Citrate, from volumes ranging from 50 μL to 200 μL. The purified plasma/serum RNA is eluted in a flexible final volume of 10 μL to 25 μL.
EXTRAClean Plasma/Serum RNA Purification Midi Kit
This utilizes a two-column method, and can purify RNA from fresh or frozen serum or plasma samples prepared from blood collected on either EDTA or Citrate, from volumes ranging from 250 μL to 1.5 mL. The first column will handle the large volume input of bodily fluids that is followed by a concentration on a mini column for a final elution of 50 μL to 100 μL.
EXTRAClean Plasma/Serum RNA Purification Maxi Kit
This kit can purify RNA from fresh or frozen serum or plasma samples prepared from blood collected on either EDTA or Citrate, from volumes ranging from 2 mL to 5 mL. The first column will handle the large volume input of bodily fluids that is followed by a concentration on a mini column for a final elution of 50 μL to 100 μL.
All sizes, including miRNA and small RNA (<200 nt)
Average Yields¥
Variable depending on specimen
† This kit is suitable for the isolation of RNA from fresh or frozen serum or plasma prepared from blood collected on either EDTA or Citrate. Plasma samples prepared from blood collected on heparin should not be used as heparin can significantly interfere with many downstream applications such as RT-PCR
¥ Please check page 5 for Average Plasma/Serum Yields and Common RNA Quantification Methods
Storage Conditions and Product Stability All solutions 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.
Human Enterovirus (HEV) TaqMan RT-PCR Detection Kits
Product Info
Document
Product Info
Overview
Detection kits for HEV
CE-IVD marked version available for in vitro diagnostic use
Available in TaqMan format for analysis
The Enteroviruses are a genus of the family Picornaviridae, and are a large and diverse group of small RNA viruses characterized by a ssRNA molecule of positive sense. To date, over 70 serotypes of Enterovirus have been isolated from man. The human enteroviruses include poliovirus, Coxsackie A and Coxsackie B viruses, enterocytopathic human orphan (ECHO) viruses and other enteroviruses. Enteroviruses infect millions of people worldwide each year, and the infection can result in a wide variety of symptoms depending on the serotype of the enterovirus. Symptoms of infection may include, but are not limited to, mild respiratory illness (common cold), hand, foot and mouth disease, Rubelliform rashes, acute hemorrhagic conjunctivitis, aseptic meningitis, myocarditis, and neonatal infections.
HEV TaqMan RT-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
HEV TaqMan RT-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 RT-PCR Master Mix (#28113) or customer supplied master mix
For research use only and NOT intended for in vitro diagnostics.
Storage Conditions and Product Stability All kit components can be stored for 1 year 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.
Purification and enrichment of intact exosomes from plasma, serum, urine, cell culture media and saliva in less than 30 minutes.
Versatile sample input ranging from 50 µL to 10 mL
Plasma/Serum Exosome Purification Mini Kit (50 µL – 1 mL Plasma/Serum)
Plasma/Serum Exosome Purification Midi Kit (1 mL – 4 mL Plasma/Serum)
Plasma/Serum Exosome Purification Maxi Kit (4 mL – 10 mL Plasma/Serum)
Exosome purification is based on Norgen’s proprietary resin separating matrix through exosomes’ surface proteins.
No precipitation reagents, overnight incubation, protease or coagulant treatments required
No time-consuming ultracentrifugation, filtration or special syringes required
Purify intact exosomes with a size ranging from 40-200 nm depending on sample input type
Purified exosomes are compatible with functional studies.
Purified exosomes are free from any RNA-binding proteins
Purified exosomes are compatible with NanoSight® or Electron Microscopy for assessing the approximate exosome size range and concentration.
Exosomal RNA can be extracted from the purified exosomes using Norgen’s Exosomal RNA Purification technology or any other RNA extraction method.
The Plasma/Serum Exosome Purification Kits provide a fast, reliable and convenient method to purify and enrich for intact exosomes from different plasma/serum sample volumes ranging from 50 µL to 10 mL. These kits also allow for the purification of intact extracellular vesicles (EVs) from different plasma/serum sample volumes, and these EVs are ready for any downstream application. The purification is based on Norgen’s proprietary resin.
These kits provide a clear advantage over other available methods since they do not require any special instrumentation, ultracentrifugation, precipitation reagents or any protease treatments. More importantly, the purified exosomes will not be contaminated with any other RNA-binding proteins that may contaminate your exosomal RNA, which is essential if studying exosomal transcripts.
NanoSight® Analysis
Exosomes enriched with Norgen’s Plasma/Serum Exosome Purification Kits can be analyzed using NanoSight® for assessing the approximate exosome size range and concentration
Exosomal RNA Analysis
To purify exosomes and isolate exosomal RNA, choose the Plasma/serum Exosome Purification and RNA isolation kits. The protocol is divided into 2 parts and an aliquot of purified exosomes can be taken for applications like NTA/TEM etc. before processing them for RNA isolation. Or you can use the Exosomal RNA Isolation Kit if you’ve already purified exosomes using a Norgen kit or another method. . Exosomal RNA isolation is based on Norgen’s proprietary resin without the need for phenol extractions or carrier RNA. This RNA is ideal for gene expression analysis using RT-qPCR, microarray, or NGS and for biomarker discovery.
Variable depending on the plasma/serum input volume
Time to Complete 10 Purifications
15 – 30 minutes
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.
Important Note This kit is suitable for the purification of exosomes from fresh or frozen serum or plasma prepared from blood collected on either EDTA or Citrate. Plasma samples prepared from blood collected on heparin should not be used as heparin can significantly interfere with many downstream applications such as RT-PCR.
