Finally, we determined the number of cells in each ectopic bone sample using DAPI staining.
Total RNA was extracted from the diagnostic bone marrow sample using the Trizol LS reagent (Invitrogen, Belgium) according to the manufacturer's recommendations.
Recently, probe capture methods for targeted enrichment have shown success in recovering degraded DNA as well as DNA from ancient bone samples using next-generation sequencing (NGS) technologies.
The extraction of DNA from the bone samples used prepared aliquots of the sample.
This evidence, however, has been questioned on several grounds: the putative non-diagnostic nature of the Dufour bladelet tool type; the proposition that other tools from the site are of Gravettian affinities; the presence of intrusive elements in the fauna; and the potential inhomogeneity of the bulk bone samples used for dating [55].
cDNA was synthesized from RNA obtained from diagnostic bone marrow samples using the Omniscript Reverse Transcription Kit (Qiagen, Chatsworth, CA, USA), according to the manufacturer's recommendations.
All parameters assumed a plate model for the trabecular bone in the derivation of the equations used for calculation, as a previous study observed for the bone samples used in this study.
Visibly, the gene expression results showed a diminished expression of all tested markers (BMP2, COL1A1, alkaline phosphatase, and osteocalcin), corroborating the abnormal functionality of osteoporotic bone samples used in this study.
Total RNA was extracted from diagnostic bone marrow or peripheral blood samples using the RNeasy Mini Kit (Qiagen) according to manufacturer's instructions.
Genomic DNA was isolated from diagnostic bone marrow or peripheral blood samples using the Qiagen DNeasy Blood and Tissue kit according to the manufacturer's instructions (Qiagen, Venio, the Netherlands).
