Details

Autor(en) / Beteiligte

• Ahlawat, Sonika
• Vasu, Mahanthi
• Choudhary, Vikas
• Arora, Reena
• Sharma, Rekha
• Mir, MA
• Singh, Manoj Kumar

Titel

Comprehensive evaluation and validation of optimal reference genes for normalization of qPCR data in different caprine tissues

Ist Teil von

• Molecular biology reports, 2024-12, Vol.51 (1), p.268-268, Article 268

Ort / Verlag

Dordrecht: Springer Netherlands

Erscheinungsjahr

2024

Quelle

MEDLINE

Beschreibungen/Notizen

• Background Quantitative real-time PCR (qPCR) is a highly reliable method for validating gene expression data in molecular studies due to its sensitivity, specificity, and efficiency. To ensure accurate qPCR results, it’s essential to normalize the expression data using stable reference genes. Methods This study aimed to identify suitable reference genes for qPCR studies in goats by evaluating 18 candidate reference genes ( ACTB, BACH1, B2M, GAPDH, HMBS, HPRT1, PGK1, PPIA, PPIB, RPLP0, RPL19, RPS9, RPS15, RPS28, SDHA, TBP, UXT , and YWHAZ ) in 10 different caprine tissues (heart, intestine, kidney, liver, lung, muscle, rumen, skin, spleen, and testis). An integrated tool called RefFinder, which incorporates various algorithms like NormFinder, GeNorm, BestKeeper, and ΔCt, was used to assess the stability of expression among these genes. Results After thorough analysis, ACTB, PPIB , and B2M emerged as the most stable reference genes, while RPL19, RPS15 , and RPS9 were found to be the least stable. The suitability of the selected internal control genes was further validated through target gene analysis, confirming their efficacy in ensuring accurate gene expression profiling in goats. Conclusion The study determined that the geometric average of ACTB, PPIB , and B2M creates an appropriate normalization factor for gene expression studies in goat tissues.