Norgen’s TaqMan 2x PCR Master Mix is a ready-to-use TaqMan 2x PCR Master Mix solution that contains a PCR internal control which can be detected by HEX/VIC channel in a real-time PCR machine. By detecting the internal control users can validate the DNA template quality, thereby preventing any false negatives in the PCR results. The user needs only to add template, target TaqMan primer/probe mix and water to set up the TaqMan real-time PCR.
PCR Control: TaqMan 2x PCR Master Mix contains PCR control primers/probe (HEX/VIC) and PCR control template. The PCR control reaction in the TaqMan 2x PCR Master Mix is optimized to not interfere with target amplification. The fluorescence of the target probe should not be HEX/VIC.
Details
Reagents Supplied – Cat # 28340
TaqMan 2x PCR Master Mix (3 Vials, 100 Reactions) – Sufficient reagent for 100 x 20 µL reactions
Storage Conditions and Product Stability Norgen’s TaqMan 2x PCR and 2x RT-PCR Master Mixes should be stored at -20ºC. For everyday use an aliquot can be stored at 4ºC for up to three months. The Master Mix is stable for multiple freeze-thaw cycles. When stored at the proper temperature this reagent is stable for at least 1 year.
Other Products
Plasma/Serum Exosome Purification and RNA Isolation Kits
Product Info
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Product Info
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.
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.
Propargyl-PEG2-amine is a crosslinker that is reactive with NHS esters, carbonyls (carboxylic acid, ketone, aldehyde). The propargyl group can participate in copper catalyzed azide-alkyne Click Chemistry reaction to form a stable triazole linkage. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
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Propargyl-PEG2-amine is a crosslinker that is reactive with NHS esters, carbonyls (carboxylic acid, ketone, aldehyde). The propargyl group can participate in copper catalyzed azide-alkyne Click Chemistry reaction to form a stable triazole linkage. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
Gel images of different ranges of library size selection. Sheared human genomic DNA was used as input.
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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.
NGS library size selection with dual clean-ups.
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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.
NGS library size selection with single clean-up for >5 kb and >10 kb libraries.
