An optimized protocol for stepwise optimization of real-time RT-PCR analysis

• Published in: Horticulture research Vol. 8; no. 1
• Main Authors: Zhao, Fangzhou, Maren, Nathan A., Kosentka, Pawel Z., Liao, Ying-Yu, Lu, Hongyan, Duduit, James R., Huang, Debao, Ashrafi, Hamid, Zhao, Tuanjie, Huerta, Alejandra I., Ranney, Thomas G., Liu, Wusheng
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 01.12.2021; Oxford University Press
• Subjects: 631/1647/2017; 631/337/572; 631/449/1659; Agriculture; Biomedical and Life Sciences; Case studies; Complementary DNA; Computer applications; Data analysis; Developmental stages; Ecology; Efficiency; Genes; Genomes; Homology; Life Sciences; Method; Nucleotide sequence; Nucleotides; Optimization; Parameter sensitivity; Plant Breeding/Biotechnology; Plant Genetics and Genomics; Plant Sciences; Polymerase chain reaction; Real time; Reverse transcription; Single-nucleotide polymorphism; Software; Soybeans

Computational tool-assisted primer design for real-time reverse transcription (RT) PCR (qPCR) analysis largely ignores the sequence similarities between sequences of homologous genes in a plant genome. It can lead to false confidence in the quality of the designed primers, which sometimes results in...