Quantifying Vimentin Gene Expression in Adult Testicular Cells

The quantity of the vimentin gene expression (V2258) in the adult testicular cells was assayed by Fluidigm (dynamic array chips). Glyceraldehyde-3-phosphate dehydrogenase was utilized as a housekeeping gene for standardization. Cultured testicular cells were selected with a micromanipulator, and lysed with a lysis buffer solution containing 1.3 μL TE buffer, 9 μL RT-PreAmp Master Mix, 0.2 μL R.T./Taq Superscript III (Invitrogen, Waltham, MA, USA), 2.5 μL 0.2× assay pool, and 5.0 μL Cells Direct 2× Reaction Mix (Invitrogen, Waltham, MA, USA). The targeted transcripts were quantified using TaqMan real-time PCR on the Biomark real-time quantitative PCR system (Fluidigm-PCR). Cells were examined in two technical repeats. Ct evaluation was analyzed by GenEx software (version 5.4.2, MultiD Analyses, Gothenburg, Sweden) [3 (link),16 (link)].