METHODS OF MEASURING GENE EXPRESSION IN FACS-SORTED CELLS

摘要

Improved methods of measuring gene expression in intracellularly immunostained FACS-sorted cell populations are provided. Exemplary methods involve fixing cells with formalin and permeabilizing with mild detergent in the presence of ribonucleoside vanadyl complex prior to FACS sorting, followed by RNA extraction in the presence of de-crosslinking agents. The resulting RNA is suitable for gene expression analysis. The method allows for analysis of the gene expression pattern specifically associated with any sortable cell population or subpopulation.

主权项

1. A method of preparing mRNA from a preparation of cells comprising:
a) adding RVC to the preparation; b) fixing the cells with formalin; c) permeabilizing the cells with mild detergent; and d) extracting mRNA in the presence of a de-crosslinking agent.