Figure 1. RNA isolation, quality control and validation of the presence of transcripts in bull spermatozoa.

(a) The cDNA was synthesized from testis RNA amplified all the primers with desired product size, whereas the cDNA synthesized from the spermatozoal RNA amplified only for spermatozoa specific primer and failed to amplify other cell specific primers indicating the purity of the spermatozoal RNA from other contaminating cells and spermatozoal genomic DNA. (b) The transcripts identified in the RNA-seq platforms were validated by qPCR. The cDNA was synthesized using Superscript-II (Invitrogen, USA) and 10 ng (based on total RNA concentration used for cDNA synthesis) of cDNA was used for qPCR reaction. The PCR products were visualized in 1.8% agarose gel electrophoresis.