Carrier RNA (cRNA) Enhances dsDNA Recovery Extracted from Small Volume Spent Embryo Culture Medium

Embryo spent culture medium has been intensively investigated, considering its promising feature for non-invasive bioanalytical techniques in in-vitro fertilization (IVF). Despite, isolating DNA from such samples is quite challenging due to its small volume. Carrier RNA is reported to exhibit DNA retrieval effects and commonly employed in various limited biological samples, but there are no reports regarding its benefit on embryo media. Therefore, we aim to evaluate the competence of cRNA on isolated DNA from embryo medium and analyzed its optimal volume as there are also no records respecting its ideal volume to obtain decent outcomes. Results showed that cRNA significantly increases DNA amounts in the cRNA treated group (p<0.001), but the D-4/D-5 medium yielded similar (p=0.684). Pearson test demonstrated no correlation between cRNA volume vs. total retrieved DNA (r=0.760, p=0.80), and Whole Genome Amplification (WGA) was shown to increase DNA in the treated group (p=0.022), but not in the untreated group (p=0.128). Additionally, electrophoresis successfully resulting in a thick and thin band of TH01 locus signifies the cRNA competence. In conclusion, our study suggests that cRNA addition is essential in embryo medium extraction as it increases initial DNA that crucial for downstream application. However, the optimal volume could not be determined in the current study since the initial amount of DNA in the medium is unknown. Obtained findings are expected to be a new input for subsequent research on DNA extraction.