title: Guanine-rich sequences inhibit proofreading DNA polymerases.
abstract: DNA polymerases with proofreading activity are important for accurate amplification of target DNA. Despite numerous efforts have been made to improve the proofreading DNA polymerases, they are more susceptible to be failed in PCR than non-proofreading DNA polymerases. Here we showed that proofreading DNA polymerases can be inhibited by certain primers. Further analysis showed that G-rich sequences such as GGGGG and GGGGHGG can cause PCR failure using proofreading DNA polymerases but not Taq DNA polymerase. The inhibitory effect of these G-rich sequences is caused by G-quadruplex and is dose dependent. G-rich inhibitory sequence-containing primers can be used in PCR at a lower concentration to amplify its target DNA fragment.
authors.affiliation: Institute of Life Sciences, Key Laboratory of Organ Development and Regeneration of Zhejiang Province, College of Life Sciences, Hangzhou Normal University, Hangzhou, Zhejiang, 310036, PR China.
authors.forename: Xiao-Jing
authors.initials: XJ
authors.lastname: Zhu
authors.affiliation: Institute of Life Sciences, Key Laboratory of Organ Development and Regeneration of Zhejiang Province, College of Life Sciences, Hangzhou Normal University, Hangzhou, Zhejiang, 310036, PR China.
authors.forename: Shuhui
authors.initials: S
authors.lastname: Sun
authors.affiliation: Institute of Life Sciences, Key Laboratory of Organ Development and Regeneration of Zhejiang Province, College of Life Sciences, Hangzhou Normal University, Hangzhou, Zhejiang, 310036, PR China.
authors.forename: Binghua
authors.initials: B
authors.lastname: Xie
authors.affiliation: Department of Anatomical Sciences and Neurobiology, University of Louisville, Louisville, KY40292, USA.
authors.forename: Xuemei
authors.initials: X
authors.lastname: Hu
authors.affiliation: Institute of Life Sciences, Key Laboratory of Organ Development and Regeneration of Zhejiang Province, College of Life Sciences, Hangzhou Normal University, Hangzhou, Zhejiang, 310036, PR China.
authors.forename: Zunyi
authors.initials: Z
authors.lastname: Zhang
authors.affiliation: Institute of Life Sciences, Key Laboratory of Organ Development and Regeneration of Zhejiang Province, College of Life Sciences, Hangzhou Normal University, Hangzhou, Zhejiang, 310036, PR China.|Department of Anatomical Sciences and Neurobiology, University of Louisville, Louisville, KY40292, USA.
authors.forename: Mengsheng
authors.initials: M
authors.lastname: Qiu
authors.affiliation: Institute of Life Sciences, Key Laboratory of Organ Development and Regeneration of Zhejiang Province, College of Life Sciences, Hangzhou Normal University, Hangzhou, Zhejiang, 310036, PR China.
authors.forename: Zhong-Min
authors.initials: ZM
authors.lastname: Dai
mesh_terms: D001483:Base Sequence; D004247:DNA / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D017931:DNA Primers / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism*; D054856:G-Quadruplexes; D020862:GC Rich Sequence / Q000235:genetics; D006147:Guanine; D016133:Polymerase Chain Reaction / Q000379:methods; D011485:Protein Binding; D019914:Taq Polymerase / Q000378:metabolism
chemical_list: D017931:DNA Primers; D006147:Guanine; D004247:DNA; D019914:Taq Polymerase; D004259:DNA-Directed DNA Polymerase