RNase Inhibitor (10,000 units) Concentration: 40 units/μL Volume: 0.25 mL Storage Conditions: Store at 2°C – 8°C Inhibits: RNases A, B, and C Supplied in: 1X PBS with 0.05% Sodium Azide.
Attogene’ s RNase Inhibitor is a protein-based ribonuclease inhibitor which noncovalently binds and inactivates a wide variety of RNases in a range of temperature (37–65°C) and pH (5.5–8.5) conditions. This product is distinct from placental RNase Inhibitor protein in that it inactivates RNases I and T1 in addition to RNases A, B and C. NA2021 is distinct from RNase Inhibitor protein in that it is less expensive, have more robust interactions with RNAses.
MADE AND FUNCTIONALLY TESTED AT ATTOGENE (MADE IN AUSTIN TEXAS – USA)
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EXTRAClean Cell Culture Media Exosome Purification and RNA Isolation Kits
Product Info
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Product Info
Overview
Ensure optimal results for sensitive applications like NGS
Up to a tenfold increase in microRNA mapping during sequencing runs to reduce costs
Purification and enrichment of intact cell culture media exosomes for functional studies
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
Pure exosomes are purified and are free-from any other RNA-binding proteins
Purification is based on EXTRAClean spin column chromatography that uses Norgen’s proprietary resin separation matrix
Norgen’s EXTRAClean Cell Culture Media Exosome Purification and RNA Isolation Kits constitute all-in-one systems for the purification of exosomes and the subsequent isolation of RNA from different cell culture media sample volumes. The purification is based on spin column chromatography that employs Norgen’s proprietary resin. The EXTRAClean columns undergo stringent processing and rigorous quality control measures to minimize contamination traces, ensuring optimal results for sensitive applications such as NGS. The kit is designed to isolate all sizes of RNA, including microRNA. The kit provides a clear advantage over other available kits in that it does not require any special instrumentation, protein precipitation reagents, extension tubes, phenol/chloroform or protease treatments. Moreover, the kits allows the user to elute into flexible elution volumes ranging from 50 μL to 100 μL. The purified RNA 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, Sequencing based applications, reverse transcription PCR, Northern blotting, RNase protection and primer extension, and expression array assays.
All sizes, including miRNA and small RNA (< 200 nt)
Elution Volume
50-100 μL
Time to Complete 10 Purifications
40-45 minutes
Average Yields
Variable depending on specimen
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. It is recommended to warm Lysis Buffer A for 20 minutes at 60°C if any salt precipitation is observed.
Isolation of exosomal RNA molecules from urine samples
Rapid and convenient spin-column protocol
Isolate inhibitor-free urinary microRNA for most downstream applications, including Small RNA Sequencing. Find out more information on Norgen’s NGS services
Optional protocol for purification of exosomal proteins for western blot analysis
This kit provides a rapid spin column procedure for the isolation of exosomal RNA from urine samples. Users can simultaneously concentrate and isolate high quality exosomal RNA, including microRNA, for use in sensitive downstream assays such as RT-PCR, qRT-PCR, NGS, microarrays and more. The protocol can be completed in under 50 minutes. Urine volumes of 1 to 10 mL can be processed easily and rapidly. All sizes of RNA are recovered at an equal rate without the need for using hazardous chemicals like phenol.
Background
Exosomes are 40 – 150 nm membrane vesicles, which are secreted by most cell types. Exosomes can be found in saliva, blood, urine, amniotic fluid and malignant ascite fluids, among other biological fluids. 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 can inform about the cell type from which the exosomes 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 urine.
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.
The series of DNA Size Selection Kits (Magnetic Beads) were developed for DNA size selection using magnetic beads. A total of 11 kits are available, with different selection ranges spanning from 50 bp to over 10 kb. The kits provide a simple and quick approach for the enrichment of a specific range of DNA fragments. The kit workflow allows double-sided or single-sided size selection for specific size cutoffs.
Gel images of different ranges of size selection. Sheared human genomic DNA was used as input.
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DNA size selection is a selective capture of DNA fragments of a specific range of size for next-generation sequencing (NGS) library preparations, PCR, ChIP assay, DNA ligations, endonuclease digestions, adapter removal, and other genomics and molecular biology applications. DNA size selection is preferred after NGS library prep in most of the cases. The NGS library preparation is related to the quality of the sequencing data. Precise NGS library size selection can increase sequencing efficiency, improve data quality, and reduce costs.
There are two types of sequencing technologies: short-read sequencing and long-read sequencing. Short-read sequencing uses DNA libraries that contain small insert DNA fragments of similar sizes, usually several hundred base pairs. The sequencing efficiency can be improved if the DNA size selection is in the right range. Cat.# 20104S and 20104L are the best kits for NGS library size selection of illumina paired-end 100 (PE100) sequencing with 100-200 bp library inserts; Cat.# 20105S and 20105L are the best kits for NGS library size selection of illumina paired-end 150 (PE150) sequencing with 150-300 bp library inserts; and Cat.# 20106S and 20106L are the best kits for NGS library size selection of illumina paired-end 300 (PE300) sequencing with 300-600 bp library inserts.
Long-read sequencing uses a large DNA fragment as input and makes very long reads. Usually, library size selection is preferred to remove smaller fragments. Cat.# 20110S and 20110L are the best kits for long-read sequencing size selection with DNA sizes >5 kb, and Cat.# 20111S and 20111L are the best kits for long-read sequencing size selection with DNA sizes >10 kb.
The magnetic beads technology uses paramagnetic particles, also known as SPRI (Solid Phase Reversible Immobilization) beads, to bind DNA reversibly and selectively. DNA fragments can be size-selected and purified by changing the properties of the magnetic beads or SPRI beads. The magnetic beads can easily separate the beads-binding DNA from the contaminants and unwanted components in the samples. The samples after DNA size selection are free of contaminants such as buffer components, enzymes, proteins, salts, dNTPs, primers, and adapters. Our proprietary magnetic beads reagents improve yield, selectivity, and reproducibility.
Specific DNA fragments at a certain length range can be purified simply using magnetic separation with different beads components, avoiding tedious and time-consuming gel extraction and column-based purification. The magnetic beads method is popular for common DNA size selection, including library size selection. The first beads-binding step, referred to as the right-side clean-up, removes large DNA fragments. The large DNA fragments are bound to the beads and are discarded. The desired DNA fragments in the supernatant are transferred to a new well, and new beads are added to the supernatant for the second beads-binding, referred to as the left-side clean-up. The double-size selected DNA fragments are eluted after ethanol rinsing.
DNA size selection with dual clean-ups.
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A single clean-up is needed for DNA size selection with large fragments. In this case, only the large DNA fragments are bound to the beads. The selected larger DNA fragments are eluted after ethanol rinsing.
DNA size selection with single clean-up for >5 kb and >10 kb DNA.
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Features of DNA size selection and library size selection
High specificity and high recovery of size selection
11 selection ranges are available, including 5 ranges for NGS library size selection
50-100 bp
100-200 bp
200-500 bp
250-350 bp: ideal for illumina PE100 sequencing
300-450 bp: ideal for illumina PE150 sequencing
450-750 bp: ideal for illumina PE300 sequencing
500-1000 bp
1-3 kb
1-5 kb
>5 kb: ideal for long-read sequencing
>10 kb: ideal for long-read sequencing
Fast and simple
20-min protocol
No gel purification required
No columns required
No centrifugation required
Efficient removal of contaminants and unwanted components