title: An improved DNA marker technique for genetic characterization using RAMP-PCR with high-GC primers.
abstract: Random amplified polymorphic DNA (RAPD) is a widely used molecular marker technique. As traditional RAPD has poor reproducibility and productivity, we previously developed an improved RAPD method (termed RAMP-PCR), which increased the reproducibility, number of bands, and efficiency of studies on polymorphism. To further develop the efficiency of this method, we used high-GC content primers for improved RAMP-PCR with DNA samples from Lonicera japonica. Comparison of amplification profiles obtained by standard RAPD primers with those obtained by regular PCR and RAMP-PCR, and high-GC primers with regular PCR and RAMP-PCR showed that the average number of bands and polymorphisms per primer gradually and significantly increased (from 6.4 to 15.0 and from 4.6 to 10.2, respectively). Cluster dendrograms showed similar results, indicating that this new method is consistent and reproducible. A total of 22 samples from different species, including plants, animals, and humans, were used for RAMP-PCR with high-GC primers. Multiple bands were successfully amplified from all samples, demonstrating that this method is a reliable technique with consistent results and may be of general interest in studies on different genera and species. We developed highly effective DNA markers, which can provide a more effective and potentially valuable approach than traditional RAPD for the genetic identification of various organisms, particularly of medicinal plants.
authors.affiliation: State Key Laboratory of Quality Research in Chinese Medicine, Macau University of Science and Technology, Macau (SAR), China.|The Research Center for Preclinical Medicine, Southwest Medical University, Luzhou, Sichuan Province, China.
authors.forename: C L
authors.initials: CL
authors.lastname: Wei
authors.affiliation: The Research Center for Preclinical Medicine, Southwest Medical University, Luzhou, Sichuan Province, China.
authors.forename: J L
authors.initials: JL
authors.lastname: Cheng
authors.affiliation: The Research Center for Preclinical Medicine, Southwest Medical University, Luzhou, Sichuan Province, China.
authors.forename: M A
authors.initials: MA
authors.lastname: Khan
authors.affiliation: The Research Center for Preclinical Medicine, Southwest Medical University, Luzhou, Sichuan Province, China.
authors.forename: L Q
authors.initials: LQ
authors.lastname: Yang
authors.affiliation: The Research Center for Preclinical Medicine, Southwest Medical University, Luzhou, Sichuan Province, China.
authors.forename: S
authors.initials: S
authors.lastname: Imani
authors.affiliation: Department of Biochemistry, School of Life Sciences & the State Key Laboratory of Medical Genetics, Central South University, Changsha City, Hunan Province, China.
authors.forename: H C
authors.initials: HC
authors.lastname: Chen
authors.affiliation: State Key Laboratory of Quality Research in Chinese Medicine, Macau University of Science and Technology, Macau (SAR), China fujunjiang@hotmail.com.|The Research Center for Preclinical Medicine, Southwest Medical University, Luzhou, Sichuan Province, China fujunjiang@hotmail.com.|Judicial Authentication Center, Southwest Medical University, Luzhou City, Sichuan Province, China fujunjiang@hotmail.com.
authors.forename: J J
authors.initials: JJ
authors.lastname: Fu
mesh_terms: D017931:DNA Primers; D018744:DNA, Plant / Q000235:genetics; D005819:Genetic Markers; D031239:Lonicera / Q000235:genetics*; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic; D019105:Random Amplified Polymorphic DNA Technique / Q000379:methods*
chemical_list: D017931:DNA Primers; D018744:DNA, Plant; D005819:Genetic Markers