The ExcelRT™ One-Step RT-PCR Kit is designed for the reverse transcription and PCR amplification of a specific target RNA from either total RNA or mRNA. The ExcelRT™ One-Step RT-PCR Kit provides the user an alternative to the lengthy two step processes (first strand generation and amplification) by using a single mixture, single tube, one step reaction. The ExcelRT™ One-Step RT-PCR Kit contains a 2X reaction premix consisting of an optimized buffer, dNTPs, Mg2+ and enzyme stabilizer, and a blend of recombinant reverse transcriptase and Taq DNA polymerase. The ExcelRT™ One-Step RT-PCR Kit allows the user to complete the RT-PCR process using a thermocycler in a single reaction setting. The ExcelRT™ One-Step RT-PCR Kit is capable of detecting even trace amounts of target RNA and ideal for target RNA amplification and analysis.
Detail
Description
The ExcelRT™ One-Step RT-PCR Kit is designed for the reverse transcription and PCR amplification of a specific target RNA from either total RNA or mRNA. The ExcelRT™ One-Step RT-PCR Kit provides the user an alternative to the lengthy two step processes (first strand generation and amplification) by using a single mixture, single tube, one step reaction. The ExcelRT™ One-Step RT-PCR Kit contains a 2X reaction premix consisting of an optimized buffer, dNTPs, Mg2+ and enzyme stabilizer, and a blend of recombinant reverse transcriptase and Taq DNA polymerase. The ExcelRT™ One-Step RT-PCR Kit allows the user to complete the RT-PCR process using a thermocycler in a single reaction setting. The ExcelRT™ One-Step RT-PCR Kit is capable of detecting even trace amounts of target RNA and ideal for target RNA amplification and analysis.
Isolate all sizes of circulating and exosomal RNA, including microRNA
Versatile urine input ranges
No phenol extractions
No carrier RNA
Bind and elute all RNA irrespective of size or GC content, without bias
Concentrate circulating RNA and exosomal RNA into flexible elution volumes
Purified RNA is suitable for a variety of downstream applications, including Small RNA Sequencing. Find out more information on Norgen’s NGS services
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 purify and concentrate high quality, high purity and inhibitor-free cell-free circulating RNA, including exosomal RNA as well as viral RNA from fresh, preserved or frozen urine samples. All components for the purification are provided in one convenient and fast kit for the easy processing of small input volumes of bodily fluids. The purified urine RNA is fully compatible with all downstream applications including PCR, qPCR, methylation-sensitive reverse transcription qPCR, reverse transcription PCR, Northern blotting, RNase protection and primer extension, expression array assays, and NGS.
Background
Recent evidence indicates that cell-free circulating RNA (cf-RNA) including exosomal RNA in urine contains valuable information for the discovery of biomarkers that can help for the early detection of certain cancer types and for monitoring the disease status as well as for the detection of any infectious pathogens. Exosomes can be found in saliva, blood, urine, amniotic fluid and malignant ascitic fluids, among other biological fluids. Evidence has been accumulating recently that these vesicles act as cellular messengers, conveying information to distant cells and tissues within the body. 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. For this reason exosomal RNAs may serve as biomarkers for various diseases including cancer. The advantage for using urine as a source for cancer biomarkers is that it can be acquired in large quantities without using invasive procedures. In addition, repeated sampling from the same individual is applicable, which facilitates longitudinal studies. There are many advantages favouring the use of urinary nucleic acid for cancer biomarker discovery over blood, tissue samples or other bodily fluids, including: (1) urine is non-infectious for HIV and less infectious for many other pathogens; (2) the profile of urinary nucleic acid is similar to that in plasma or serum but with a lower concentration; (3) Nucleic acid purification from urine is technically much easier because of its low protein concentration (1000-fold lower than blood).
Urine Cell-Free Circulating RNA Purification Mini Kit
For sample volumes ranging from 2 mL to 10 mL. The first column will handle the large volume input of urine that is followed by a concentration on a mini column for a final elution of 50 µL to 100 µL.
For sample volumes ranging from 10 mL to 30 mL. The first column will handle the large volume input of urine that is followed by a concentration on a mini column for a final elution of 50 µL to 100 µL
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
*Please check page 6 of the product insert for average yields and common RNA quantification methods.
Storage Conditions and Product Stability All buffers should be kept tightly sealed and stored at room temperature. These kits are 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.
t-Boc-aminooxy-PEG6-propargyl is a bifunctional linker molecule. The propargyl group can react with azide-bearing compounds or biomolecules via copper catalyzed azide-alkyne Click Chemistry to yield a stable triazole linkage. The protected aminooxy can be deprotected under mild acidic conditions and then can react with an aldehyde.. The hydrophilic PEG spacer increases solubility in aqueous media. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
Document
t-Boc-aminooxy-PEG6-propargyl is a bifunctional linker molecule. The propargyl group can react with azide-bearing compounds or biomolecules via copper catalyzed azide-alkyne Click Chemistry to yield a stable triazole linkage. The protected aminooxy can be deprotected under mild acidic conditions and then can react with an aldehyde.. The hydrophilic PEG spacer increases solubility in aqueous media. Reagent grade, for research purpose. Please contact us for GMP-grade inquiries.
Produced by reverse osmosis, ion-exchange, electro-deionization, ultrafiltration, UV treatment (UVC & 185 nm), 0.1 μm membrane filtration
Nuclease-free, protease-free, endotoxin-free
Suitable for molecular biology applications (qPCR, NGS etc.)
Suitable for cell culture applications
Produced in an ISO certified GMP laboratory
Water system is routinely monitored for compliance with QC standards
The Norgen water purification system has been designed in conjunction with experts from Millipore Sigma. The Ultrapure NFW is purified in a two-stage process. The first stage, to produce type II water, includes pre-treatment with activated charcoal and polyphosphate, this water then undergoes reverse osmosis, electro-deionization and ion exchange to remove contaminants as well as UV treatment for disinfection. The Pure water then flows to a second system in a clean room with an additional 4 layers of purification. After passing through an additional ion exchange column, the water is then subjected to Ultrafiltration to remove particulates, bacteria, & ionic contaminants down to trace level, the water is then treated with UV light (182 nm wavelength) to break down organic matter. Lastly a second Ultrafilter for the production of pyrogen-, nuclease- and bacteria-free water at 18.2 MΩ. This water is of utmost purity and suitable for direct use in molecular biology applications such as qPCR and NGS, as well as cell culture and other applications.
Product Specifications
pH
n/a (highly pure water does not contain enough ions for an exact pH determination.)
Recommended Storage
Room Temperature
Grade
Ultrapure (ASTM Type I)
Biological Activity
DNase-, RNase-, Protease-, Endotoxin- Free
TOCs
< 50 ppb
Resistivity
> 18 MΩ-cm
Treatment
Not DEPC-Treated
Quantity
100 mL, 500 mL, 1000 mL
Purification Methods
Activated charcoal, polyphosphate, reverse osmosis, electro-deionization, ion exchange, UV, ultrafiltration
Shipping Conditions
Room Temperature
Details
Applications For use in any molecular biology application
Storage Conditions Norgen’s Nuclease-Free Water should be stored at room temperature. Storage at +4°C or -20°C is optional.
Precautions and Disclaimers This product is designed for research purposes only. It is not intended for human or diagnostic use.
