Gel documentation imaging system

The total DNA from the sperm cells was isolated using the QIAamp DNA Mini kit (QIAGEN) as per the manufacturer’s instruction: https://www.qiagen.com. Three microliters of the extracted DNA and 2 μl of DNA gel loading dye (6×) were mixed then loaded in a 1.5% agarose gel to determine the integrity of DNA, and electrophoresis was carried out at 100 V/cm for 30 min. The photograph was examined below UV light by a gel documentation imaging system (Bio-Rad, USA). The extracted DNA was quantified using the NanoDrop Lite Spectrophotometer (Thermo Scientific).

Total RNA samples were harvested using the TriReagent (Invitrogen, USA) following the manufacturer’s protocols. First strand cDNA was then synthesized using the Reverse Transcription System (Promega) in a thermal cycler (MJ Research Inc. USA). Forward (5′- AAT GAA TTC TG ATG TCT TCC GTA TTC GAT G -3′) and reverse (5′-AAT CTC GAG C TCA ATA CCC CCA GTC GGT GT -3′) primers were used to amplify the NDV NP gene using 30 PCR cycles. The conditions were 94°C for 30 s, 55°C for 1 min, and 72°C for 1.5 min, followed by a final extension step of 72°C for 7 min. The resulting PCR product was electrophoresed, stained with ethidium bromide and viewed with a Gel Documentation Imaging System (BioRad, USA).

The GeNei^TMmini-submarine gel system was used to investigate the multiplex of gene expression in linoelaidic acid-treated MCF-7 cells. The primers of all genes were supplied by Bioserve (Telangana, India). Briefly, 5 μl of PCR product was mixed with 1 μl sample loading dye and 5 μl of DNA size standard 3000. Run using GeNei^TMmini-submarine gel system (co. name). The amplified fragments were separated according to their respective size by a mini-submarine gel system. Results were analyzed using the Gel Documentation Imaging System (Bio-Rad, Model No. 1708275).