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Automating RNA Isolation


Large scale gene expression screening projects require reliable high-throughput methods to isolate high quality RNA from a large number of samples. To meet this growing need, Ambion developed the RNAqueous-96 Kit. This kit, which utilizes an RNA-binding glass-fiber filter, provides high yields of intact RNA in a 96-well plate format. An optional on-the-filter DNase treatment can be performed to ensure removal of genomic DNA for RT-PCR applications. Because the kit is compatible with commonly used 96-well plate vacuum manifolds, the procedure is readily automated for use with high throughput liquid handling systems.

Recently, Ambion, in collaboration with PerkinElmer Life Sciences, developed an automation protocol for the RNAqueous-96 Kit using the Packard MultiPROBE II HT Liquid Handling System. This method reproducibly yields high quality total RNA from both adherent and suspended mammalian cultured cells.


Reproducible Isolation of High Quality RNA
To demonstrate the high quality of RNA that can be obtained using the RNAqueous-96 Kit on a robotic platform, RNA from multiple HeLa S3 and K562 cell samples was isolated using the method described in "RNAqueous-96 Automation Protocol". Briefly, HeLa S3 (adherent) and K562 (suspended) cells were transferred to a deep well 96-well plate at 3.75 x 105 cells/well. HeLa cells were incubated for 2 days prior to RNA isolation, whereas K562 cells were processed immediately. The resulting RNA was assessed for both yield and qua
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