The DirectBlood Genotyping Kit is compatible with a wide range of assays: Here we are detecting a SNP in the Factor V gene (rs6025; G1691A), which is connected with a higher risk for venous thrombosis, with a fluorescent hydrolysis probe assay.
Homozygous wild type blood samples will show a strong signal and amplification plot (blue curve) in channel 2 (Cy5) and none or a very low signal
(red curve) in the other channel 1 (ROX).
Heterozygous blood samples will give an intermediate signal and amplification plots with very similar intensities in both channels 1 and 2 (blue and red curve). Homozygous mutant blood samples, channel 1 (ROX) will show a strong signal and amplification plot (red curve) and channel 2 (Cy5) none or a very low signal and amplification plot (blue curve). So the signals are reversed on homozygous mutant samples.
Here we are detecting the same SNP in the Factor V gene (rs6025; G1691A), which is connected with a higher risk for venous thrombosis, with a fluorescent hybridization probe assay.
Three blood samples with known genotype were used. All the genotypes can clearly be identified. The blue curve shows the specific wild type (homozygous G;G) peak at 61°C. The red curve shows the specific mutant (homozygous A;A) peak at 53°C. A heterozygous case (A;G) will result in both melting curve peaks, respectively at the same specific temperature for wild type and mutant (orange curve).
DirectBlood Genotyping PCR Kit is a novelty on the market. It allows genotyping in real-time and without any previous DNA isolation. You simply save time and money: directly use EDTA blood without any extraction steps.
The DirectBlood Genotyping PCR Kit is optimized to function with a wide range of different SNPs to detect possible nucleotide variations. It has been successfully tested with a variety of hydrolysis probes and hybridization probes.
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.
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| Kit Specifications | |
| Plasma/Serum Input (Cat. 57400) | 50 μL – 1 mL |
| Plasma/Serum Input (Cat. 57500) | 1 mL – 4 mL |
| Plasma/Serum Input (Cat. 57600) | 4 mL – 10 mL |
| Size of Exosomes Purified | 40 nm – 150 nm |
| Elution Volume | 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.
| Component | Cat. 57400 (50 preps) | Cat. 57500 (25 preps) | Cat. 57600 (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 |
| Mini Filter Spin Columns inserted into 2 mL tubes | 50 | 25 | 15 |
| Product Insert | 1 | 1 | 1 |
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Library size selection is an enrichment of a specific range of library sizes for NGS library preparations. 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, or SPRI (Solid Phase Reversible Immobilization) beads, is well used for the purification of DNA due to their reversible DNA binding. The NGS library can be size-selected by the magnetic beads or SPRI beads. The properties of the magnetic beads can be changed for a specific range of DNA binding. The contaminants and other unwanted components in the libraries can also be removed during size selection.
Specific ranges of NGS libraries can be selected using magnetic beads with different buffer compositions. The first DNA-beads binding step, also called the right-side clean-up, removes large NGS library fragments. The large NGS library fragments that bind to the beads are discarded with the beads pellet. The desired NGS library fragments in the supernatant are transferred to a new well, and new beads are added to the supernatant for the second beads-DNA binding, also called the left-side clean-up. After the rinsing step, the NGS library fragments with the dual selection are eluted in water or an appropriate buffer. The magnetic beads method has great advantages over time-consuming column purification and tedious gel-based purification.
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Library size selection for long-read sequencing only requires a single clean-up. In this case, only the large library fragments are bound to the beads, while other small library fragments are discarded with the supernatant. The selected larger library fragments are eluted in water or an appropriate buffer after the rinsing step.
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