Development of novel SCAR markers for genetic characterization of Lonicera japonica from high GC-RAMP-PCR and DNA cloning

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. Th...

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Bibliographic Details
Published in:Genetics and molecular research 2016-01, Vol.15 (2)
Main Authors: Cheng, J L, Li, J, Qiu, Y M, Wei, C L, Yang, L Q, Fu, J J
Format: Article
Language:English
Subjects:
Cloning, Molecular - methods
DNA Primers - chemistry
DNA Primers - genetics
GC Rich Sequence
Genetic Markers
Lonicera - genetics
Lonicera japonica
Random Amplified Polymorphic DNA Technique - methods
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Summary: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.
ISSN:1676-5680
1676-5680
DOI:10.4238/gmr.15027737