non methylated control dna  (Zymo Research)

Journal: Scientific Reports

Article Title: Epigenetic modulation of VEGF-A/VEGFR2 pathway genes in OC/TME axis driving genetic upregulation and tumor plasticity

doi: 10.1038/s41598-025-04483-6

Figure Lengend Snippet: Representative agarose gel images showing MSP product analysis of VEGF-A, VEGF-C, and VEGFR2 genes in ovarian tumors, benign samples and respective peritoneal fluid (PF) DNA samples. For each case, universally methylated genomic DNA (M+) served as a positive control for methylated alleles, while universally unmethylated genomic DNA (U+) served as a positive control for unmethylated alleles. In the VEGF-A gene, the methylated (M) band size is 205 bp, and the unmethylated (U) band size is 207 bp. In the VEGF-C gene, the methylated (M) band size is 131 bp, and the unmethylated (U) band size is 129 bp. In the VEGFR2 gene, the methylated (M) band size is 193 bp, and the unmethylated (U) band size is 186 bp. The PCR product in M-lane indicates the methylated allele, the PCR product in U-lane indicates the unmethylated allele and bands in both lanes represent partial methylation. Samples S01-S05 are carcinoma cases, B01-B03 are benign cases and PF1-PF3 are peritoneal fluid cases.

Article Snippet: The PCR protocol included 95 °C initial denaturation for 10 min, 40 cycles of 95 °C denaturation for 15 s, annealing at the specified temperature for 1 min (Supplementary Table 1), 60 °C primer extension for 1 min, and a final 72 °C extension for 5 min. Universal methylated and unmethylated human DNA (Zymo Research, USA) served as positive control, while distilled water without DNA was the negative control.

Techniques: Agarose Gel Electrophoresis, Methylation, Positive Control