title: Enhancement of PCR amplification of moderate GC-containing and highly GC-rich DNA sequences.
abstract: PCR is a commonly used and highly efficient technique in biomolecular laboratories for specific amplification of DNA. However, successful DNA amplification can be very time consuming and troublesome because many factors influence PCR efficiency. Especially GC-rich DNA complicates amplification because of generation of secondary structures that hinder denaturation and primer annealing. We investigated the impact of previously recommended additives such as dimethylsulfoxide (DMSO), magnesium chloride (MgCl2), bovine serum albumin (BSA), or formamide. Furthermore, we tested company-specific substances as Q-Solution, High GC Enhancer, and Hi-Spec; various actively promoted polymerases as well as different PCR conditions for their positive effects on DNA amplification of templates with moderate and extremely high CG-content. We found considerable differences of specificity and quantity of product between different terms. In this article, we introduce conditions for optimized PCR to help resolve problems amplifying moderate to high GC-rich templates.
authors.affiliation: Institute of Legal Medicine, Jena University Hospital-Friedrich Schiller University Jena, Fürstengraben 23, 07743 Jena, Germany. Juliane.strien@med.uni-jena.de
authors.forename: Juliane
authors.initials: J
authors.lastname: Strien
authors.forename: Juliane
authors.initials: J
authors.lastname: Sanft
authors.forename: Gita
authors.initials: G
authors.lastname: Mall
mesh_terms: D001482:Base Composition; D004247:DNA / Q000032:analysis / Q000737:chemistry / Q000235:genetics; D017931:DNA Primers; D004121:Dimethyl Sulfoxide / Q000737:chemistry; D005559:Formamides / Q000737:chemistry; D020862:GC Rich Sequence; D006801:Humans; D015636:Magnesium Chloride / Q000737:chemistry; D016133:Polymerase Chain Reaction / Q000379:methods*; D012710:Serum Albumin, Bovine / Q000737:chemistry
chemical_list: D017931:DNA Primers; D005559:Formamides; D015636:Magnesium Chloride; D012710:Serum Albumin, Bovine; C031066:formamide; D004247:DNA; D004121:Dimethyl Sulfoxide