PAXgene Bone Marrow RNA Kit

The supernatants resulting from the first two spin steps of the PAXgene Bone Marrow RNA Kit purification procedure may still contain biologically hazardous material. We recommend classifying the tube-processed supernatant of the kit as both a biological and chemical waste. Supernatants can be decontaminated by autoclaving and disposed of as chemical waste.  Disposal should be done in accordance with local, state and federal regulations.

CAUTION: Do not add bleach or acidic solutions directly to the sample preparation waste.

The PAXgene Bone Marrow RNA Tubes should be frozen at –20°C and –70°C in a wire rack, not in an expanded polystyrene foam tray.

If the PAXgene Bone Marrow RNA Tubes are to be stored at temperatures lower than –20ºC, freeze them first for 24 h at –20ºC, then transfer them to –70ºC.

Currently available data shows stabilization of cellular RNA for several months at –20°C or –70°C. For more information from ongoing studies evaluating stability for longer time periods, please contact QIAGEN Technical Services.

Bone marrow samples are extremely heterogeneous, consisting of varying amounts of cells, tissue, and solid matter. RNA yields are highly donor-dependent and can vary greatly from sample to sample as well as between replicates from the same donor due to sample-intrinsic heterogeneity.

In a study of 295 bone marrow samples, yields of RNA isolated from 2 mL human bone marrow were between 2.0 and 44.7 µg (5-95% quantiles, mean = 15.8 µg, median = 11.0 µg).

No. The 260/230 nm absorbance ratio is not specified for the PAXgene Bone Marrow RNA extraction kit. In contrast to a reduced 260/280 ratio, which results from a protein contamination of the eluted RNA and can lead to inhibitory effects in down stream applications, there is no negative impact of a reduced 260/230 ratio on any down stream application reported to date and therefore it is not necessary to specify this.

In addition, because the elution buffer contains very low concentrations of salt that absorb at wavelengths between 220 and 230 nm, it is necessary to zero the photometer properly. Do this by preparing a solution for zeroing the photometer which contains a similar salt concentration as the sample to be measured by using the same volumes. Add the same volume of Buffer BMR5 as the volume of RNA sample to be diluted into a fresh volume of the buffer in which you will measure the absorption (we recommend 10 mM Tris Cl pH 7.5 buffer for this purpose).