DECALCIFICATION SOLUTION WITH PRESERVATION OF RNA

摘要

The present invention is directed to methods of decalcification and tissue sample preparation that allows for the reproducible quantitative analysis of gene expression in hard tissue samples like bone, mineralizing cartilage and tendon, dentin, cementum and/or enamel that are too hard to section effectively using conventional means.

主权项

1. A method of decalcification of hard tissues like bone, mineralizing cartilage and tendon, dentin, cementum and enamel for sectioning without degrading the ribonucleic acids (RNA) contained therein, comprising the steps of:
A. obtaining a hard tissue sample and placing it in an RNA preservation solution; B. removing said hard tissue sample from the RNA preservation solution and placing it in a liquid decalcification medium comprising an RNA preservation solution and a decalcification agent, wherein said decalcification agent is selected from the group consisting of tetrasodium-EDTA or trisodium-EDTA wherein said decalcification agent begins to remove calcium from mineral deposits in said hard tissue sample; C. stirring the mixture of step B said until said hard tissue sample softens for sectioning; and D. rinsing the softened hard tissue sample of step C in an RNA preservation solution to remove any remaining decalcification agent.