The 96 Well Total RNA Kit is designed for the automated high throughput isolation of total cellular RNA from animal and human cells. Up to 1 x 10⁶ animal or human cells can be processed in each well of the 96 Well RNA plate. High quality total cellular RNA is uniformly extracted in each well of the plate. This kit is ideal for quantitative analytic applications.

Principle

The 96 Well Total RNA Kits use reversible binding properties of HiBind^® matrix, a new silica-based, time saving spin technology material. Combined with the speed of mini column spin technology or vacuum manifold, multiple samples can be processed at the same time. Initially the sample is lysed under denaturing buffer conditions so to inactivate RNases, thus protecting intact RNA from degradation. After adjusting the buffer conditions, samples are loaded to the HiBind^® RNA Plate, where a brief centrifugation or vacuum, forces the samples through the plate allowing the RNA to bind to the Hibind^® matrix. After two wash steps, purified RNA can be eluted with DEPC treated RNase-free water.

Procedure

Cells are lysed by the addition of 150 µl TRK Lysis Buffer to each well on the 96 well tissue culture plate. The pate is subsequently agitated vigorously for 30 seconds. Equal volumes of 70% ethanol are added to the sample to precipitate RNA enabling it to bind to the column matrix. Samples are applied to the wells of he HiBind^® RNA plate. Plates are centrifuged for 5 minutes at 5000 x g at room temperature. Once spin step is completed the plates are washed Wash Buffer I and plates can be treated with an on column DNase treatment by directly pipetting DNase on the surface of the HiBind^® RNA resin in each column (optional). Plates are allowed to incubate at room temperature(25-30°C) for 15 minutes. After incubation plates are washed with RNA Wash Buffer I and RNA Wash Buffer II (diluted with ethanol). An additional spin or vacuum step is applied to the HiBind^® RNA plate and RNA is then eluted with DEPC treated water.

Applications

RNA purified using the Total RNA method can be used in applications such as :

• cDNA Libraries
• RT-PCR
• Northern Blot