myo-Inositol Assay Kit

Description

The EzRNA™ RNA Capping System is a user-friendly product for post-transcriptional RNA modification. Both Vaccinia Capping Enzyme and 2’-O-Methyltransferase are included in the package, which are able to perform in a single reaction. The Vaccinia Capping Enzyme attach 7-methylguanylate cap (m7Gppp, Cap-0) to the 5′ end of RNA to form m7Gppp5’N-RNA (Cap-0 RNA). The 2′-O-methyltransferase utilizes Cap-0 RNA as a substrate, employing S-adenosine methionine (SAM) as a methyl donor to methylate 2′ -OH of the first nucleotide at the 5′ end of Cap-0 RNA, resulting in the formation of Cap-1 RNA.

Features

• 2’-O-Methyltransferase included for Cap-1 RNA
• High capping efficiency
• High stability
• RNase inhibitor is included to enhance the stability of capping reaction

Application

• Generation of 5’Cap-0 (m7Gppp) and Cap-1 (m7GpppNm-) RNA by enzymatic reaction
• mRNA synthesis for in vitro translation
• Gene expression studies
• mRNA vaccine development and therapeutics

Storage

-20°C for 24 months

The EzRNA™ RNA Capping System is a user-friendly product for post-transcriptional RNA modification. Both Vaccinia Capping Enzyme and 2’-O-Methyltransferase are included in the package, which are able to perform in a single reaction. The Vaccinia Capping Enzyme attach 7-methylguanylate cap (m7Gppp, Cap-0) to the 5′ end of RNA to form m7Gppp5’N-RNA (Cap-0 RNA). The 2′-O-methyltransferase utilizes Cap-0 RNA as a substrate, employing S-adenosine methionine (SAM) as a methyl donor to methylate 2′ -OH of the first nucleotide at the 5′ end of Cap-0 RNA, resulting in the formation of Cap-1 RNA.

Overview

• Isolate all sizes of RNA, including microRNA, irrespective of size or GC content, without bias
• Versatile sample input ranges
• Isolate all sizes of free-circulating RNA, including microRNA​
• Bind and elute all RNA irrespective of size or GC content, without bias
• The purified exosomal RNA is free from any circulating RNA-binding proteins
• No phenol extractions, Proteinase K treatment nor carrier RNA required
• No time-consuming ultracentrifugation, filtration nor special syringes are required
• No precipitation reagents nor overnight incubation required
• Concentrate isolated exosomal RNA and free-circulating RNA into a flexible elution volume ranging from 50 µL to 100 µL
• Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix

These kits provide a fast, reliable and convenient method to sequentially isolate and concentrate exosomal RNA as well as Free-Circulating RNA from different urine 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 since they do not require any special instrumentation, protein precipitation reagents, extension tubes, phenol/chloroform or protease treatments. Moreover, the kits allow the user to elute into a flexible elution volume ranging from 50 µL to 100 µL. The RNA isolated from the purified exosomes is free from any protein-bound circulating RNA and is of the highest integrity. Moreover, the free-circulating, protein-bound, RNA is free from any exosomal RNA. 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.

Urine Exosome and Free-Circulating RNA Isolation Mini Kit

For sample volumes ranging from 250 µL to 1 mL.

Urine Exosome and Free-Circulating RNA Isolation Midi Kit

For sample volumes ranging from 2 mL to 10 mL.

Urine Exosome and Free-Circulating RNA Isolation Maxi Kit

For sample volumes ranging from 11 mL to 30 mL.

Details

Supporting Data

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Kit Specifications
Minimum Urine Input	11 mL
Maximum Urine Input	30 mL
Size of RNA Purified	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 the specimen

*Please check page 5 of the product insert for the average yields and the common RNA quantification methods.

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 shipping. It is recommended to warm Lysis Buffer A for 20 minutes at 60°C if any salt precipitation is observed.

Important Note
Urine samples stored at -70°C, -20°C or at 4°C will develop some precipitation due to the aggregation of some of the highly abundant proteins in urine. Eliminating these precipitates using centrifugation or filtration may cause the loss of exosomes. Furthermore, these precipitates may affect the quality of the purified nucleic acid. We recommend the use of Norgen’s Urine Preservative when collecting urine samples, which is designed for the preservation of nucleic acids and proteins in fresh urine samples at ambient temperatures. The components of the Urine Preservative allow samples to be stored for over 2 years at room temperature with no detected degradation of urine DNA, RNA or proteins. Norgen’s Urine Preservative is available as a liquid format in Norgen’s Urine Preservative Single Dose Ampules, as well as in a dried format in Norgen’s Urine Collection and Preservation Tubes.

Component	Cat. 59200 (50 preps)	Cat. 59300 (25 preps)	Cat. 59400 (15 preps)
Slurry E	12.5 mL	12.5 mL	12.5 mL
ExoC Buffer	8 mL	30 mL	50 mL
ExoR Buffer	12 mL	12 mL	12 mL
Lysis Buffer A	2 x 20 mL	2 x 20 mL	2 x 20 mL
Lysis Additive B	2 mL	2 mL	2 mL
Wash Solution A	2 x 18 mL	18 mL	18 mL
Elution Solution A	2 x 6 mL	6 mL	6 mL
Mini Filter Spin Columns	50	25	15
Mini Spin Columns	100	50	30
Collection Tubes	100	50	30
Elution Tubes (1.7 mL)	100	50	30
Product Insert	1	1	1

Description

FluoroDye™ DNA Fluorescent Loading Dye is a ready-to-use 6X DNA loading dye designed for fast qualitative electrophoresis analysis. Containing sensitive fluorescent dye with high specific affinity towards double stranded DNA (dsDNA), the FluoroDye™ Fluorescent DNA Loading Dye has negligible background and renders destaining process unnecessary. The FluoroDye™ DNA Fluorescent Loading Dye allows the user to immediately visualize electrophoresis result upon completion or to monitor the electrophoresis in real time. FluoroDye™ DNA Fluorescent Loading Dye is compatible with both the conventional UV gel-illuminating system as well as the less harmful long wavelength blue light illumination system. FluoroDye™ emission as bound to dsDNA is 522 nm, while its excitation peaks are at 270, 370 and 497 nm.

Features:

• Excellent for premix with DNA samples
• Sensitivity: 0.14 ng (DNA)
• A safer alternative to EtBr
• Compatibility: suitable to blue or UV light
• Increased cloning efficiency (blue light)

Composition 

FluoroDye™ DNA Fluorescent Loading Dye is stored in 6X concentration in 60% glycerol and buffered with Tris-HCl and EDTA, containing Bromophenol blue, Xylene cyanol FF and Orange G as tracking dyes.

Storage

Protected from light
 -20°C for 24 months

FluoroDye™ DNA Fluorescent Loading Dye is a ready-to-use 6X DNA loading dye designed for fast qualitative electrophoresis analysis. Containing sensitive fluorescent dye with high specific affinity towards double stranded DNA (dsDNA), the FluoroDye™ Fluorescent DNA Loading Dye has negligible background and renders destaining process unnecessary. The FluoroDye™ DNA Fluorescent Loading Dye allows the user to immediately visualize electrophoresis result upon completion or to monitor the electrophoresis in real time. FluoroDye™ DNA Fluorescent Loading Dye is compatible with both the conventional UV gel-illuminating system as well as the less harmful long wavelength blue light illumination system. FluoroDye™ emission as bound to dsDNA is 522 nm, while its excitation peaks are at 270, 370 and 497 nm.