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Biotech Trading Partners |
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Biotech Trading Partners |
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QnE-RNA Kit
Catalog# BTP-100 2 X 96 Purifications
The QnE-RNA Kit is designed for the high-throughput purification of total cellular RNA from cultured cells in a 96-well plate format. The kit can also be used to isolate total cellular RNA from plasma or serum, or to cleanup up to 96 RNA samples at a time. No phenol-chloroform extraction or precipitation steps are involved. The sample addition and washing steps are performed using any 96-well compatible vacuum manifold, while the final elution of the RNA product is performed using a table-top centrifuge.
Cells are lysed under denaturing conditions using a lysis buffer containing the denaturant guanidine isothiocyanate. Ethanol is next added to the samples, which are then added to the filter plates. This step facilitates the binding of RNA to the filter plate’s silica membrane matrix. Under these conditions the RNA binds to the membrane while other contaminants are washed through. The plate is then washed to further remove protein, buffer components and other contaminants using two ethanol-containing wash buffers and the final RNA product is eluted in RNase-free water. The final RNA product is high quality total cellular RNA that can be used directly for quantitative RT-PCR and other downstream applications. A protocol to treat the final RNA product with DNase I is also included, but most applications will not require its use since very little DNA contaminants are found in the final RNA product. Small RNAs (<200 nucleotides) such as tRNA are not efficiently isolated using this kit.
The QnE-RNA Kit 96-well silica membrane plates have a standard Society for Biomolecular Screening (SBS) footprint and are compatible with a variety of automated liquid handling workstations, all 96-well-compatible vacuum manifolds and most, if not all, 96-well plate-compatible rotors. In fact, these plates are directly compatible with QIAGEN’s RNeasy 96 kit, Invitrogen’s PureLink 96 RNA Purification kit, Ambion’s RNAqueous 96 kit, and Promega’s SV 96 Total RNA Isolation Kit. All of these kits utilize a similar guanidine-based lysis buffer, followed by RNA binding to a silica membrane filter matrix, washing of the membrane and then sample elution.
The RNA extraction protocol will take 30-45 min. to perform, however two vacuum manifolds in parallel can be used to process two 96-well plates in nearly the same amount of time.
In our experience, the quality and quantity of cellular RNA isolated from the QnE-RNA Kit (and replacement plates), the linearity of RNA content with cell number and the interwell variation are superior to those found in all other comparable 96-well RNA purification kits. |