title: Rapid and simple method of qPCR primer design.
abstract: Quantitative real-time polymerase chain reaction (qPCR) is a powerful tool for analysis and quantification of gene expression. It is advantageous compared to traditional gel-based method of PCR, as gene expression can be visualized "real-time" using a computer. In qPCR, a reporter dye system is used which intercalates with DNA's region of interest and detects DNA amplification. Some of the popular reporter systems used in qPCR are the following: Molecular Beacon(®), SYBR Green(®), and Taqman(®). However, success of qPCR depends on the optimal primers used. Some of the considerations for primer design are the following: GC content, primer self-dimer, or secondary structure formation. Freely available software could be used for ideal qPCR primer design. Here we have shown how to use some freely available web-based software programs (such as Primerquest(®), Unafold(®), and Beacon designer(®)) to design qPCR primers.
authors.affiliation: Treasure Coast High School, 1000 SW Darwin Boulevard, Port St. Lucie, FL, 34953, USA.
authors.forename: Brenda
authors.initials: B
authors.lastname: Thornton
authors.forename: Chhandak
authors.initials: C
authors.lastname: Basu
mesh_terms: D017931:DNA Primers / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012984:Software
chemical_list: D017931:DNA Primers