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.

OverView

T7 RNA Polymerase catalyzes the formation of RNA from a DNA template in the 5’-3’ direction and is commonly used in in vitro transcription (IVT) applications. The enzyme is T7 promoter specific, requires Mg²⁺ as a cofactor and can use modified nucleotides for the synthesis. The T7 RNA Polymerase requires a double-stranded DNA template and can produce full-length RNA transcripts.

Key Features

• T7 promoter-specific RNA polymerase
• Available in standard glycerol-based formulation as well as lyophilization-friendly formulation without glycerol.

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Suggested Applications

• In vitro transcription of RNA
• Molecular diagnostics (NASBA and other)

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Properties

Quality Control

ArcticZymes is dedicated to the quality of our products. T7 RNA Polymerase is manufactured at our ISO 13485 certified facility in Norway.‍

T7 RNA Polymerase catalyzes the formation of RNA from a DNA template in the 5’-3’ direction and is commonly used in in vitro transcription (IVT) applications. The enzyme is T7 promoter specific, requires Mg2+ as a cofactor and can use modified nucleotides for the synthesis. The T7 RNA Polymerase requires a double-stranded DNA template and can produce full-length RNA transcripts.