Description :

Introduction

The EZgene^TM Plant RNA Midiprep Kit provides a convenient and rapid method for isolating total RNA from a variety of plant samples. The kit introduces a homogenization column to efficiently remove cell debris and simultaneously homogenize the lysate. In combination with ezBind RNA spin column, high quality RNA can be extracted from as much as 500 mg tissue. Typical yields are shown in Table I. The EZgene^TM Plant RNA Kits are ideal for processing multiple plant samples in less than one hour. The need for organic extractions is eliminated, making total RNA isolation fast, safe, and reliable. Purified RNA has OD_260/280 ratios of 1.8-2.0 and is suitable for the following applications.

•  RT-PCR
•  Northern Analysis
•  Differential display
•  Poly A+ RNA selection

Storage and Stability

All components of the EZgene^TM Plant RNA Kit should be stored at 22-25 ^oC. Under these conditions, RNA has successfully been purified and used for RT-PCR after 24 months of storage. Under cool ambient conditions, a precipitate may form in the Buffer LY. In case of such an event, heat the bottle at 37 ^oC to dissolve.  Store Buffer LY at room temperature.

Binding Capacity

Each ezBind RNA column can bind approximately 100 μg RNA. Using greater than 500 mg plant tissue usually will not dramatically improve yields and sometimes has adverse effects.

Kit Contents

**Buffer LY contains a chaotropic salt. Use gloves and protective eyeware when handling this solution.

*DNase I are not supplied. They could be purchased from Biomiga.

Before Starting

Please take a few minutes to read this booklet thoroughly to become familiar with the protocol. Prepare all materials required before starting to minimize RNA degradation.

•  Whenever working with RNA, always wear latex gloves to minimize RNase contamination. Change gloves frequently. Use only clean RNase-free disposable plastic pipette tips when using the supplied reagents.

•  During the procedure work carefully but quickly.
•  Under cool ambient conditions, crystals may form in Buffer LY. This is normal and the bottle may be warmed to dissolve the salt.

•  β-mercaptoethanol (β-mercaptoethanol) is the key in denaturing endogenous RNases and must be added to an aliquot of Buffer LY before use. Add 10 μL of ß-mercaptoethanol per 1 mL of Buffer LY. This mixture can be stored for 1 week at room temperature.

Trouble Shooting Guide

Oprating Protocol: