Digestible and Resistant Starch Assay Kit

Overview

• Purification and enrichment of intact plasma/serum exosomes for functional studies
• Isolate all sizes of RNA, including microRNA, irrespective of size or GC content, without bias
• Versatile plasma/serum input volume
• No phenol extractions, Proteinase K treatment, nor carrier RNA required
• No time-consuming ultracentrifugation, filtration nor special syringes required
• No precipitation reagents, nor overnight incubation required
• Compatible with plasma/serum from most species
• Pure exosomes are purified and are free from any other RNA-binding proteins
• Purification is based on spin column chromatography that uses Norgen’s proprietary resin
• The purified RNA is suitable for a variety of downstream applications, including Small RNA Sequencing. Find out more information on Norgen’s NGS services

Norgen’s Plasma/Serum Exosome and RNA Isolation Kits constitute all-in-one systems for the purification of exosomes and the sequential isolation of exosomal RNA from different plasma/serum sample volumes. The purification is based on spin column chromatography that employs Norgen’s proprietary resin. These kits are designed to isolate all sizes of RNA, including microRNA, irrespective of size or GC content, without bias. These kits provide a clear advantage over other available kits in that they do not require any special instrumentation, protein precipitation reagents, extension tubes, phenol/chloroform or protease treatments. The RNA isolated from the purified exosomes is free from any protein-bound circulating RNA and is of the highest integrity. The purified RNA 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.

Norgen’s Plasma/Serum Exosome and RNA Isolation Mini Kit

For sample volumes ranging from 50 µL to 1 mL. This kit allows the user to elute into a flexible elution volume ranging from 50 µL to 100 µL.

Norgen’s Plasma/Serum Exosome and RNA Isolation Midi Kit

For sample volumes ranging from 1 mL to 4 mL. This kit allows the user to elute into a flexible elution volume ranging from 50 µL to 100 µL.

Norgen’s Plasma/Serum Exosome and RNA Isolation Maxi Kit

For sample volumes ranging from 4 mL to 10 mL. This kit allows the user to elute into a flexible elution volume ranging from 50 µL to 100 µL.

Details

Supporting Data

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Kit Specifications
Minimum Plasma/Serum Input	4 mL
Maximum Plasma/Serum Input	10 mL
Size of Exosomes Purified	40 nm – 150 nm
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

*Please check page 4 of the product insert for the average yields and the 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.

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.

Component	Cat. 58300 (50 preps)	Cat. 58500 (25 preps)	Cat. 58600 (15 preps)
Slurry E	12.5 mL	12.5 mL	12.5 mL
ExoC Buffer	8 mL	8 mL	2 x 8 mL
ExoR Buffer	12 mL	12 mL	12 mL
Lysis Buffer A	20 mL	20 mL	20 mL
Lysis Additive B	2 mL	2 mL	2 mL
Wash Solution A	38 mL	18 mL	18 mL
Elution Solution A	6 mL	6 mL	6 mL
Mini Filter Spin Columns	50	25	15
Mini Spin Columns	50	25	15
Collection Tubes	50	25	15
Elution Tubes (1.7 mL)	100	50	30
Product Insert	1	1	1

Description

African Swine Fever Virus (ASFV) is a widespread disease which infects members of the pig family(Suidae). Anumberoftick species are believed to be the vector for the disease,as well as being transmitted by raw pork and pig excrement [1]. After firstly being identified in Kenya in 1921, ASFV became endemic in sub-Saharan Africa, with regular outbreaks being reported across Europe, Asia and South America throughout the century [2]. More recently the virus was introduced in Georgia and spread throughout the region, as well as mass outbreaks occurring in China in 2018 [3].
ASFVistheonlymemberoftheAsfaridaefamily.ItisalargeenvelopeddoublestrandedDNA virus of icosahedral morphology with an average diameter of 200nm and isolates contain genomes between 170-190Kbp encoding for up to 167 open reading frames [2]. The morphology of ASFV consist of several concentric domains. An inner core contains the nucleoid coated with a thick protein layered core shell, which is surrounded by an inner lipid envelope , all of which is encompassed by the capsid [2]. ASFV begins its replication cycle in the nucleus of infected cells before moving to the cytoplasm where the majority of the replication takes place [2]. Gene transcription is highly regulated, with distinct classes of mRNA identified to accumulate at early, intermediate and late transcripts of the virus [2]. The disease induces acute haemorrhagic disease within its hosts, causing high fevers and skin haemorrhages, with death often occurring within ten days of clinical symptoms appearing [4].

References: 1: The Centre for Food Security and Public Health (2015), African Swine Fever. 2: Galindo, I. and Alonso, C., 2017. African swine fever virus: a review. Viruses, 9(5), p.103. 3: Zhou, X., Li, N., Luo, Y., Liu, Y., Miao, F., Chen, T., Zhang, S., Cao, P., Li, X., Tian, K. and Qiu, H.J., 2018. Emergence of African swine fever in China, 2018. Transboundary and emerging diseases, 65(6), pp.1482-1484. 4: Gallardo, C., Ademun, A.R., Nieto, R., Nantima, N., Arias, M., Martín, E., Pelayo, V. and Bishop, R.P., 2011. Genotyping of African swine fever virus (ASFV) isolates associated with disease outbreaks in Uganda in 2007. African Journal of biotechnology, 10(17), pp.3488-3497.

Exceptional value for money
Rapid detection of all clinically relevant subtypes
Positive copy number standard curve for quantification
Highly specific detection profile
High priming efficiency
Broad dynamic detection range (>6 logs)
Sensitive to < 100 copies of target Accurate controls to confirm findings

DBCO-C5-PEG12-acid is an analog of DBCO-Acid with PEG linker and a DBCO group. DBCO group is a highly reactive cycloalkyne which is commonly used for copper-free Click Chemistry reactions. The hydrophilic PEG chain allows for increased water solubility of compounds in aqueous media. The terminal carboxylic acid can react with primary amine groups in the presence of activators (e.g. HATU) to form a stable amide bond. Reagent grade, for research use only.

DBCO-C5-PEG12-acid is an analog of DBCO-Acid with PEG linker and a DBCO group. DBCO group is a highly reactive cycloalkyne which is commonly used for copper-free Click Chemistry reactions. The hydrophilic PEG chain allows for increased water solubility of compounds in aqueous media. The terminal carboxylic acid can react with primary amine groups in the presence of activators (e.g. HATU) to form a stable amide bond. Reagent grade, for research use only.