Takara NucleoSpin 96 RNA Blood \4 x 96 Preps

The NucleoSpin 96 RNA Blood kit is (4 x 96 preps) is designed for purifying RNA from whole blood on 96-well plates. Samples are lysed in chaotropic salts, which immediately inactivate RNases present in virtually all biological materials and create the binding conditions favoring adsorption of RNA to the silica membrane on the plate. Contaminating DNA, which is also bound to the silica membrane, is removed by the recombinant DNase solution. Samples can be processed using either vacuum or centrifugation.

Effective DNA removal. Total RNA was isolated manually under vacuum from 400-µl whole blood samples (EDTA) according to the standard protocol for NucleoSpin 96 RNA Blood, with and without an on-column rDNase treatment. A PCR for human elongation factor EF-1a was performed to determine the amount of residual genomic DNA in the eluted RNA using a LightCycler system and gene-specific primers designed to yield a 469-bp product. The figure shows the corresponding amplification curves of the 0.01 ng–100 ng DNA standards (dark blue lines), the NTC (negative control; green), and replicates of the blood sample 1 (1-1, 1-2), with DNase treatment (+ DNase; red lines) and without DNase treatment (-DNase; light blue lines). In total, blood samples from six individual donors were analyzed by PCR (data not shown). Samples treated with rDNase (+DNase) show an average Cp of 31.8. Samples purified without rDNase (-DNase) treatment show an average Cp of 25.9. Omitting the on-column DNA digestion results in an approximately 64 times higher amount of residual gDNA.

Consistent RNA quality from fresh or frozen blood samples.NucleoSpin 96 RNA Blood was used to isolate RNA in triplicate from 400-µl fresh or frozen blood (EDTA) samples obtained from six different donors under vacuum. Frozen sample aliquots had been stored at –20°C. RNA quality was determined by spectrophotometric measurement of absorption at 260 nm and 280 nm.

Consistent RNA yield from fresh or frozen blood samples.NucleoSpin 96 RNA Blood was used to isolate RNA in triplicate from 400-µl fresh or frozen blood (EDTA) samples obtained from six different donors under vacuum. Frozen sample aliquots had been stored at –20°C. RNA yield was determined by spectrophotometric measurement of absorption at 260 nm and 280 nm.