title: Development of novel SCAR markers for genetic characterization of Lonicera japonica from high GC-RAMP-PCR and DNA cloning.
abstract: Sequence-characterized amplified region (SCAR) markers were further developed from high-GC primer RAMP-PCR-amplified fragments from Lonicera japonica DNA by molecular cloning. The four DNA fragments from three high-GC primers (FY-27, FY-28, and FY-29) were successfully cloned into a pGM-T vector. The positive clones were sequenced; their names, sizes, and GenBank numbers were JYHGC1-1, 345 bp, KJ620024; YJHGC2-1, 388 bp, KJ620025; JYHGC7-2, 1036 bp, KJ620026; and JYHGC6-2, 715 bp, KJ620027, respectively. Four novel SCAR markers were developed by designing specific primers, optimizing conditions, and PCR validation. The developed SCAR markers were used for the genetic authentication of L. japonica from its substitutes. This technique provides another means of developing DNA markers for the characterization and authentication of various organisms including medicinal plants and their substitutes.
authors.affiliation: Key Laboratory of Epigenetics and Oncology, The Research Center for Preclinical Medicine, Sichuan Medical University, Luzhou, Sichuan, China.
authors.forename: J L
authors.initials: JL
authors.lastname: Cheng
authors.affiliation: Key Laboratory of Epigenetics and Oncology, The Research Center for Preclinical Medicine, Sichuan Medical University, Luzhou, Sichuan, China.
authors.forename: J
authors.initials: J
authors.lastname: Li
authors.affiliation: Key Laboratory of Epigenetics and Oncology, The Research Center for Preclinical Medicine, Sichuan Medical University, Luzhou, Sichuan, China.|Maternal and Child Health Care Hospital of Zigong, Zigong, Sichuan, China.
authors.forename: Y M
authors.initials: YM
authors.lastname: Qiu
authors.affiliation: Key Laboratory of Epigenetics and Oncology, The Research Center for Preclinical Medicine, Sichuan Medical University, Luzhou, Sichuan, China.|State Key Laboratory of Quality Research in Chinese Medicine, Macau University of Science and Technology, Macau, China.
authors.forename: C L
authors.initials: CL
authors.lastname: Wei
authors.affiliation: Key Laboratory of Epigenetics and Oncology, The Research Center for Preclinical Medicine, Sichuan Medical University, Luzhou, Sichuan, China.
authors.forename: L Q
authors.initials: LQ
authors.lastname: Yang
authors.affiliation: Key Laboratory of Epigenetics and Oncology, The Research Center for Preclinical Medicine, Sichuan Medical University, Luzhou, Sichuan, China.|State Key Laboratory of Quality Research in Chinese Medicine, Macau University of Science and Technology, Macau, China.|Judicial Authentication Center, Sichuan Medical University, Luzhou City, Sichuan, China.
authors.forename: J J
authors.initials: JJ
authors.lastname: Fu
mesh_terms: D003001:Cloning, Molecular / Q000379:methods*; D017931:DNA Primers / Q000737:chemistry / Q000235:genetics; D020862:GC Rich Sequence; D005819:Genetic Markers; D031239:Lonicera / Q000235:genetics*; D019105:Random Amplified Polymorphic DNA Technique / Q000379:methods*
chemical_list: D017931:DNA Primers; D005819:Genetic Markers