title: Pseudogene-free amplification of HPRT1 in quantitative reverse transcriptase polymerase chain reaction.
abstract: Quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) provides a powerful tool for precise gene expression analysis. The accuracy of the results highly depends on careful selection of a reference gene for data normalization. HPRT1 (hypoxanthine phosphoribosyl transferase 1) is a frequently used housekeeping gene for normalizing relative expression values. However, the existence of processed pseudogenes for HPRT1 might interfere with reliable results obtained in qRT-PCR due to amplification of unintended products. Here, we designed a primer pair for pseudogene-free amplification of HPRT1 in qRT-PCR. We demonstrate that this primer pair specifically amplified HPRT1 messenger RNA (mRNA) sequence while avoiding coamplification of the pseudogenes.
authors.affiliation: Molecular and Cell Biology Research Center (MCBRC), Faculty of Medicine, Mazandaran University of Medical Sciences, Sari, Mazandaran, Iran; Department of Immunology, Faculty of Medicine, Mazandaran University of Medical Sciences, Sari, Mazandaran, Iran.
authors.forename: Reza
authors.initials: R
authors.lastname: Valadan
authors.affiliation: Molecular and Cell Biology Research Center (MCBRC), Faculty of Medicine, Mazandaran University of Medical Sciences, Sari, Mazandaran, Iran.
authors.forename: Omolbanin
authors.initials: O
authors.lastname: Amjadi
authors.affiliation: Molecular and Cell Biology Research Center (MCBRC), Faculty of Medicine, Mazandaran University of Medical Sciences, Sari, Mazandaran, Iran; Department of Immunology, Faculty of Medicine, Mazandaran University of Medical Sciences, Sari, Mazandaran, Iran.
authors.forename: Mohsen
authors.initials: M
authors.lastname: Tehrani
authors.affiliation: Molecular and Cell Biology Research Center (MCBRC), Faculty of Medicine, Mazandaran University of Medical Sciences, Sari, Mazandaran, Iran; Department of Immunology, Faculty of Medicine, Mazandaran University of Medical Sciences, Sari, Mazandaran, Iran. Electronic address: rafiei1710@gmail.com.
authors.forename: Alireza
authors.initials: A
authors.lastname: Rafiei
authors.affiliation: Molecular and Cell Biology Research Center (MCBRC), Faculty of Medicine, Mazandaran University of Medical Sciences, Sari, Mazandaran, Iran.
authors.forename: Akbar
authors.initials: A
authors.lastname: Hedayatizadeh-Omran
authors.affiliation: Molecular and Cell Biology Research Center (MCBRC), Faculty of Medicine, Mazandaran University of Medical Sciences, Sari, Mazandaran, Iran.
authors.forename: Reza
authors.initials: R
authors.lastname: Alizadeh-Navaei
mesh_terms: D000818:Animals; D002460:Cell Line; D017931:DNA Primers / Q000737:chemistry* / Q000235:genetics*; D006801:Humans; D007041:Hypoxanthine Phosphoribosyltransferase / Q000737:chemistry / Q000235:genetics*; D012333:RNA, Messenger / Q000737:chemistry / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*
chemical_list: D017931:DNA Primers; D012333:RNA, Messenger; D007041:Hypoxanthine Phosphoribosyltransferase
keywords: HPRT1; Pseudogenes; Reference gene; qRT-PCR