Convenient optimized on-column DNase treatment using Norgen’s RNA Purification Kits
Also includes protocol for digestion in-solution followed by RNA Clean-Up
Guaranteed RNase-Free
Includes Enzyme Incubation Buffer
Cat. 25710 contains one vial of 1,600 units and Cat. 25720 contains 4 vials (1,600 units/vial)
Norgen’s RNA purification kits isolate total RNA with minimal amounts of genomic DNA contamination. However, for some sensitive downstream applications, it may be desirable to remove all traces of residual DNA. Norgen’s RNase-free DNAse I Kit, with Enzyme Incubation Buffer, can be used for optional on-column DNase digestion with any of Norgen’s RNA purification kits. Alternatively, after isolating total RNA using one of Norgen’s RNA purification kits, the RNA elution can be treated with this DNase I. The RNA can then be purified from the DNase using Norgen’s RNA Clean-Up and Concentration Kit (Cat# 23600), and the RNA can then be used in downstream applications.
Details
Each RNase-Free DNase I Kit is supplied complete with sufficient enzyme and enzyme incubation buffer for 50 or 200 reactions.
Storage Conditions The DNase I provided is in lyophilized form. It is stable for at least 3 months if stored at room temperature. However, it is recommended to store the DNase I vial at 2 – 8ºC (or below) upon receipt to maintain stability beyond 3 months. Buffer DR and Enzyme Incubation Buffer can be stored at room temperature. After reconstitution with Buffer DR (see product manual), the DNase I should be stored at -20ºC. All reagents should remain stable for at least 1 year in their unopened containers at the appropriate storage temperature.
The Kit is designed for purification of total RNA, including miRNA and other small RNA molecules (18nt), from cultured cells and various animal and human tissues, including difficult-to-lyse tissues samples. Alternatively, a miRNA-enriched fraction and a total RNA (>200nt) fraction can be purified separately. This Kit combines phenol/guanidine-based lysis of samples and silica membrane–based purification of total RNA. MagZol Reagent, included in the kits, is a monophasic solution of phenol and guanidine thiocyanate, designed to facilitate lysis of tissues, to inhibit RNases, and also to remove most of the cellular DNA and proteins from the lysate by organic extraction.
Details
Specifications
Features
Specifications
Main Functions
Isolation miRNA and other small RNA molecules(18nt), from cultured cells and various animal and human tissues, using MagZol reagent and column
Applications
RT-PCR, Northern Blot, poly A+purification, nucleic acid protection and in vitro translation
Hipure silica gel column is based on glass fiber filter membrane with high binding force. Under the condition of high concentration of ionizing agent (such asguanidine hydrochloride or guanidine isothiocyanate), the filter membrane can adsorb nucleic acid through hydrogen bond and electrostatic, while protein andother impurities are not adsorbed and removed. The filter membrane adsorbed with nucleic acid is washed to remove the residual protein and salt. Finally, the nucleic acid adsorbed on the filter membrane can be washed out with low salt buffer (such as buffer TE) or water. The obtained nucleic acid has high purity and can be directly used in various downstream experiments.
This kit combines acid/guanidine (MagZol) extraction technology with glass fiber filter membrane purification, which can improve the extraction effect of complex samples and samples with low RNA content. After the sample is treated with MagZol reagent and chloroform, the supernatant is added with ethanol to provide appropriate binding conditions, then transferred to the purification column and centrifuged. Macromolecular RNA can be efficiently bound to the membrane. Collect the filtrate containing small RNA, add more ethanol to adjust the binding capacity of small RNA, the pollutants can be efficiently washed away by second cleaning. Finally, the purified RNA was eluted by RNase free water.
Advantages
High quality – one step RNA extraction reagent combined with silica gel column can obtain the highest concentration
Fast – several samples can be extracted in 40 minutes
High applicability – samples including animals, plants, bacteria, cells, etc.
High concentration – efficiently remove macromolecular RNA, enrich small RNA and improve sensitivity
Kit Contents
Contents
R431002
R431003
Purification Times
50 Preps
250 Preps
HiPure RNA Mini Columns
100
2 x 250
2ml Collection Tubes
100
2 x 250
MagZol Reagent
60 ml
270 ml
Buffer RWC
20 ml
80 ml
Buffer RW2*
20 ml
2 x 50 ml
RNase Free Water
10 ml
30 ml
Storage and Stability
MagZol Reagent should be stored at 2-8°C upon arrival. However, short-term storage (up to 24 weeks) at room temperature (15-25°C) does not affect its performance. The remaining kit components can be stored at room temperature (15-25°C) and are stable for at least 18 months under these conditions.
Document
The Kit is designed for purification of total RNA, including miRNA and other small RNA molecules (18nt), from cultured cells and various animal and human tissues, including difficult-to-lyse tissues samples. Alternatively, a miRNA-enriched fraction and a total RNA (>200nt) fraction can be purified separately. This Kit combines phenol/guanidine-based lysis of samples and silica membrane–based purification of total RNA. MagZol Reagent, included in the kits, is a monophasic solution of phenol and guanidine thiocyanate, designed to facilitate lysis of tissues, to inhibit RNases, and also to remove most of the cellular DNA and proteins from the lysate by organic extraction.
CE-IVD marked version available for in vitro diagnostic use
Available in TaqMan format for analysis
Norovirus is a single-stranded, non-enveloped RNA virus belonging to the Caliciviridae family. Norovirus is considered the major causative agent of non-bacterial gastroenteritis in the world. The main channel of transmittance is via contaminated food or water, as well as human-to-human contact. Most Noroviruses infecting humans belong to genogroup I and II (particularly genotype 4). Norovirus is very stable in the environment and is resistant to some surface disinfectants such as alcohols and detergents. Moreover, it is very difficult to culture Norovirus in vitro, making its identification by standard microbiological assays challenging.
NoV TaqMan RT-PCR Kit, 100 reactions
Ready to use format, including Master Mix for the target and PCR control to monitor for PCR inhibition and validate the quality
Specific Primer and Probe mix for the pathogen/virus/viroid of interest
Primer and Probe mix
Positive and negative control to confirm the integrity of the kit reagents
NoV TaqMan RT-PCR Probe/Primer Set and Controls, 100 reactions
Specific Primer/Probe mix and Positive Control for the pathogen/virus/viroid of interest
Nuclease-free water
Can be used together with Norgen’s RT-PCR Master Mix (#28113) or customer supplied master mix
Storage Conditions and Product Stability All kit components can be stored for 1 year after the date of production without showing any reduction in performance.
All kit components should be stored at -20°C upon arrival. Repeated thawing and freezing (> 2 x) of the Master Mix and Positive Control should be avoided, as this may affect the performance of the assay. If the reagents are to be used only intermittently, they should be frozen in aliquots.