High throughput RNA extraction from Mus musculus tissue using a Tissue RNA kit and the Bead Ruptor 12

The extraction and isolation of RNA is an integral part of downstream analyses such as RT-PCR, RT-qPCR, Northern blotting, and cDNA library construction. The importance of using pure, intact RNA for these processes is well documented and a critical part of downstream analysis success. It is well known that RNA is sensitive to degradation due to mechanical shear, temperature, storage conditions and freeze-thawing. Furthermore, RNA is highly susceptible to RNAse degradation following release of nucleases during the tissue disaggregation process. Thus, proper sample handling during the homogenization process is crucial when performing an RNA based assay.

Bead-based homogenization upstream of RNA extraction can help streamline the preparation of tissue when compared to manual methods. Herein, we demonstrate the Omni Bead Ruptor 12 bead mill homogenizer in a multi-tissue RNA workflow comparing bead based sample prep to manual cryomilling techniques. RNA integrity and yield were measured downstream and used to drive comparison between sample preparation methods.

Table 1: Average RNA yield for and RIN value for each tissue type processed on the Omni Bead Ruptor 12.