The Plant Total RNA Kit provides a convenient and rapid method for the isolation of total RNA from a variety of plant samples. The system is efficient enough to allow isolation of total RNA from as little as 10 mg of plant tissue, while containing interfering compounds like polysaccharides, starches and phenolics. Plant Total RNA Kits are ideal for processing multiple samples in less than one hour.

Principle

The Plant RNA Kit includes homogenizing columns to simultaneously and efficiently remove cell wall debris and homogenize the sample. The method involves a simple and rapid precipitation step for the removal polysaccharides, starches and phenolic compounds commonly found in plant tissues. In combination with HiBind^® RNA spin-columns, this procedure permits purification of high quality RNA from as much as 100 mg tissue, which is suitable for a number of downstream applications. The use of mini-columns facilitate the binding, washing, and elution steps thus enabling multiple samples to be simultaneously processed. For more information on columns, click Flexible Multi-Format Columns.

Procedure

Collect approximately 100 mg of finely ground plant material in a microfuge tube and immediately add 500 µl Buffer RB containing the appropriate amount of ß-mercaptoethanol and incubate. After incubation transfer lysate into a Homogenization Spin Column and centrifuge at 13,000 x g for 5 minutes at room temperature. After centrifugation, discard the homogenization column and add 0.5 volume ethanol and mix by inverting tube. Apply the sample, including any precipitate to a HiBind^® RNA spin column and centrifuge at 10,000 x g for 30 sec at room temperature. If highly pure RNA is required add DNase I digestion reaction mix from the RNase Free DNase I Digestion Set (Catalog Number E1091-01/02) directly onto the surface of the HiBind® RNA resin and incubate. Once incubation is complete the column is subjected to a variety of Wash Steps prior to elution from the column with DEPC-treated water.

Applications

RNA purified using the Plant RNA kit is suitable for the following applications :

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

Gel analysis of RT-PCR. Total RNA were isolated from five leaves sample of
potato (lane 1), tomato (lane 2), barley (lane 3), pea (lane 4), and wheat (lane 5).