( Rapidly isolate...)High-Throughput Isolation of Total RNA

Rapidly isolate total RNA from 96 samples

Karen Dolter Sylvia Norman Jeff Braman
Stratagene

The StrataPrep 96 total RNA purification kit provides a convenient method for simultaneous isolation of total RNA from up to 96 cultured cell samples. Because RNA is bound to a solid support, there is no need for organic extraction and alcohol precipitation. The resulting RNA is high quality, with no detectable DNA contamination, and is ready for molecular beacon and conventional RT-PCR applications.

Isolating total RNA from many samples by traditional methods can be time-consuming and tedious. The StrataPrep method is efficient and allows processing of a wide range of sample sizes, from 10 cells to 5 x 10⁵ cells per well.

Protocol

Fig.1

An overview of the StrataPrep 96 protocol is shown in Figure 1. After cultured cells are disrupted by guanidine thiocyanate, lysates are applied to a silica-based fiber matrix within the 96-well plate. RNA is immobilized on the fiber matrix, and contaminants are removed while avoiding organic extractions and ethanol precipitation. The sample is treated with DNase directly on the fiber matrix and pure RNA is recovered by eluting with a small volume of buffer.

The standard protocol makes use of a vacuum manifold and a tabletop centrifuge. If a vacuum manifold is not available, an alternate protocol, which uses the centrifuge alone, may be used (additional materials required).

Isolation of RNA from Various Cell Lines

Table 1: Average Yields of RNA from Various Cells (g)

* Numbers in parenthesis indicate the number of isolations performed.

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