A normalisationof targeted genes using the appropriate reference genes is necessary for reliable quantitative reverse transcription polymerase chain reaction (qRT-PCR) experiments. This study aimed to investigate the appropriate reference genes and the numbers of the genes required for qRT-PCR in canine pulmonary arteries. Pulmonary arteries were collected from twenty-three dogs at necropsy. Candidate reference genesincluding ribosomal protein L32 (RPL32), ribosomal protein L13a (RPL13A), ribosomal protein S18 (RPS18), TATA box binding protein (TBP), beta-2-microglobulin (B2M), ribosomal protein S5 (RPS5), ribosomal protein S19 (RPS19) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) from pulmonary arteries were examined by qRT-PCR and analysed using four major algorithms, including geNorm, NormFinder, the comparative delta-CTmethod, and RefFinder. The optimal number of genes was analysed using geNorm. The ranking of the appropriate reference genes from all algorithms was evaluated by RefFinder. The appropriate reference genes for the normalisation of the targeted gene in canine pulmonary arteries are RPS19 and RPL32.
Tangmahakul, Nattawan; Techangamsuwan, Somporn; and Surachetpong, Sirilak Disatian
"Selection of the appropriate reference genes for relative quantitative reverse transcription polymerase chain reaction (qRT-PCR) in canine pulmonary arteries,"
The Thai Journal of Veterinary Medicine: Vol. 49:
4, Article 1.
Available at: https://digital.car.chula.ac.th/tjvm/vol49/iss4/1