Introduction

MagZol Reagent is a reagent system for the isolation of total RNA from cells and tissues. The reagent, a single-phase solution consisting of phenol and guanidine isothiocyanate, is modification of the single-step RNA isolation method developed by Chomczynski and Sacchi. The sample is homogenized and lysed in MagZol Reagent which maintains the integrity of the RNA, while disrupting and denaturing endogenous RNases and other cellular components. Extraction of the lysate with chloroform further denatures proteins and separates the mixture into an organic and an aqueous phase. RNA remains exclusively in the aqueous phase, and is subsequently recovered by isopropanol.

This method is suitable for small quantities of tissue (<100mg) and cells (<5 X10⁶), and large quantities of tissue (up to 1g) and cells (<10⁸), of human, animal, plant, or bacterial origin. The simplicity of the MagZol Reagent method allows simultaneous processing of a large number of samples. The entire procedure can be completed in one hour. Total RNA prepared in this manner can be used for Northern blot analysis, dot blot hybridization, poly(A) + selection, in vitro translation, RNase protection assay, and molecular cloning. For use in amplification by thermal cycling, treatment of the isolated RNA with RNase-free DNase I is recommended when the two amplimers lie within a single exon.

Details

Specifications

Advantages

• Flexible – sample amount can be adjusted according to the demand
• Cost performance -the most economical nucleic acid extraction technology

Storage and Stability

MagZol Reagent should be stored at 2-8°C upon arrival and is stable for at least 24 months under the condition. However, short-term storage (up to 12 weeks) at room temperature (15-25°C) does not affect its performance.

MagZol Reagent is a reagent system for the isolation of total RNA from cells and tissues. The reagent, a single-phase solution consisting of phenol and guanidine isothiocyanate, is modification of the single-step RNA isolation method developed by Chomczynski and Sacchi. The sample is homogenized and lysed in MagZol Reagent which maintains the integrity of the RNA, while disrupting and denaturing endogenous RNases and other cellular components. Extraction of the lysate with chloroform further denatures proteins and separates the mixture into an organic and an aqueous phase. RNA remains exclusively in the aqueous phase, and is subsequently recovered by isopropanol.

Overview

• Excellent separation and purification of cytoplasmic and nuclear RNA
• Convenient and fast spin column format
• High quality and yield of RNA
• Isolate full diversity of RNA (including microRNA) without phenol
• Purified RNA is ready for any application including RT-PCR, qRT-PCR, RNA-Seq, arrays and more
• Cytoplasmic RNA is free of DNA and ready for direct use in RT-PCR/qRT-PCR
• Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix

This kit provides a rapid method for the isolation and purification of both cytoplasmic and nuclear RNA from cultured animal cells and small tissue samples. The kit can be used to isolate all sizes of RNA from the cytoplasmic and nuclear RNA fractions, including all small RNA species without any requirement for phenol.   Included in the kit are sufficient reagents to perform either 50 cytoplasmic RNA preparations or 25 cytoplasmic and 25 nuclear RNA preparations.   Ten samples can be processed in approximately 45 minutes.  This kit is also available in a 100 prep size.    

Background

In certain circumstances it is desirable to be able to isolate fractionated RNA as opposed to total RNA. For example, it may be preferable to isolate only mature, cytoplasmic RNA for some studies on expression profiling. Alternatively it may be desirable to isolate nuclear RNA in order to investigate and study pre-processed (non-spliced) RNA. Furthermore, this kit can be used to isolate RNA for downstream applications where it is necessary to avoid DNA contamination, since the cytoplasmic fraction has been shown to be free of all traces of genomic DNA. 

Details

Supporting Data

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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.

Introduction

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

Principle

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

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.

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.