Optimization and Primers Screening of SRAP-PCR System in Symplocos paniculata(PDF)
《北方园艺》[ISSN:1001-0009/CN:23-1247/S]
- Issue:
- 2018年24
- Page:
- 27-34
- Research Field:
- Publishing date:
Info
- Title:
- Optimization and Primers Screening of SRAP-PCR System in Symplocos paniculata
- Author(s):
- AN Baiyi1; YU Huiying1; WU Shuang1; LIU Xiaojia1; SUN Xiaogang1; ZHANG Yan2
- (1.College of Horticulture,Jilin Agricultural University,Changchun,Jilin 130118;2.Agro-biotechnology Research Institute,Jilin Academy of Agricultural Sciences,Changchun,Jilin 130124)
- Keywords:
- Symplocos paniculata; SRAP; system optimization; primers screening
- PACS:
- -
- DOI:
- 10.11937/bfyy.20181074
- Abstract:
- In order to establish and optimize SRAP-PCR reaction system in Symplocos paniculata (Thunb.) Miq.,the fresh leaves of Symplocos paniculata were used as experimental materials,the combination of single factor test and orthogonal test were used to optimize SRAP-PCR reaction system with 5 factors,and SRAP primer combinations were screened.The results showed that,the influence of each factor on the result of SRAP-PCR amplification was quite different,and the descending order was DNA＞dNTPs＞Mg2+＞primer＞Taq DNA polymerase.A suitable SRAP-PCR system for Symplocos paniculata was that total 10 μL reaction system containing 075 U Taq DNA polymerase,01 mmol·L-1 dNTPs,2 mmol·L-1 Mg2+,04 μmol·L-1 primer and 80 ng DNA.A total of 25 polymorphic SRAP primer combinations were screened from 176 SRAP primer combinations.PCR products were stably amplified among plants by different primers,which indicated that the SRAP-PCR system was suitable for molecular markers research of Symplocos paniculata.
Last Update: 2019-01-16