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Biotech Trading Partners |
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QnE-BLOOD RNA KIT
Catalog# BTP-500 2 X 96 Purifications
The QnE-Blood RNA kit is designed for the high-throughput purification of total cellular RNA from whole blood in a 96-well plate format. 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 lysis of red blood cells (RBCs) and the final elution of the RNA product are performed using a table-top centrifuge.
The red blood cells (RBCs) in whole blood are first selectively lysed and the while blood cells are pelleted by centrifugation. The white blood cells are then lysed under denaturing conditions using a 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-Blood 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.
The RNA extraction protocol will take approximately 1.5-2 hr. 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-Blood RNA Kit, the linearity of RNA content with blood volume and the interwell variation are superior to those found in all other comparable 96-well RNA purification kits. |