Optimization of Chinese fir SRAP-PCR system

doi:10.3969/j.issn.1000-2006.2014.01.003

WANG Xinmin,ZHENG Renhua,CHEN Jinhui,ZHAO Yaqi,XU Yang,WANG Ying,SHI Jisen

JOURNAL OF NANJING FORESTRY UNIVERSITY ›› 2014, Vol. 38 ›› Issue (01) : 15-20.

PDF(2712803 KB)

JOURNAL OF NANJING FORESTRY UNIVERSITY ›› 2014, Vol. 38 ›› Issue (01) : 15-20. DOI: 10.3969/j.issn.1000-2006.2014.01.003

Optimization of Chinese fir SRAP-PCR system

WANG Xinmin¹,ZHENG Renhua²,CHEN Jinhui¹,ZHAO Yaqi¹,XU Yang¹,WANG Ying¹,SHI Jisen^1*

Author information +

History +

Abstract

In order to optimize SRAP-PCR reaction system for Chinese fir,the orthogonal design L₁₆(4⁵)was considered with different concentrations of Mg²⁺, dNTPs, primer, Taq DNA polymerase and DNA template in SRAP-PCR reaction system for Chinese fir. The results of PCR were evaluated by visual and variance analysis,then the concentrations of Mg²⁺,dNTPs,primer were rectified with single-factor design. Finally,the results showed that the best reaction system was the combination in 20 μL reaction solution,containing of Mg²⁺ 2.25 mmol/L,dNTPs 0.15 mmol/L,primer 0.4 μmol/L,Taq DNA polymerase1.5 μmol/min and DNA template 60 ng,2 μL10×PCR Buffer and ddH₂O to 20 μL. The first optimal annealing temperature was 35 ℃,and the second step was 53 ℃. The optimized reaction system of Chinese fir described above can get clear and stable bands.

Cite this article

EndNote

Ris (Procite)

Bibtex

Download Citations

WANG Xinmin,ZHENG Renhua,CHEN Jinhui,ZHAO Yaqi,XU Yang,WANG Ying,SHI Jisen. Optimization of Chinese fir SRAP-PCR system[J]. JOURNAL OF NANJING FORESTRY UNIVERSITY. 2014, 38(01): 15-20 https://doi.org/10.3969/j.issn.1000-2006.2014.01.003

References

[1] 于永福.杉科植物的起源、演化及其分布[J].植物分类学报,1995,33(4):362-389.Yu Y F.Origin,evolution and distribution of the Taxodiaceae[J].Acta Phytotaxonomica Sinica,1995,33(4):362-389.
[2] 侯振宏,张小全,徐德应,等.杉木人工林生物量和生产力研究[J].中国农学通报,2009,25(5):97-103.Hou Z H,Zhang X Q,Xu D Y,et al.Study on biomass and productivity of Chinese fir plantation[J].Chinese Agricultural Science Bulletin,2009,25(5):97-103.
[3] 郑仁华,施季森.福建省杉木良种繁育现状与对策[J]. 林业科技开发,2004,18(2):3-7.Zheng R H,Shi J S.The present situation and the counter measures of Cunninghamia lanceolata breeding in Fujian province[J].China Forestry Science and Technology,2004,18(2):3-7.
[4] 郑仁华,施季森,翁玉榛,等.福建省杉木育种战略研究[J].林业科技开发,2008,22(2):1-6.Zheng R H,Shi J S,Weng Y Z,et al.The reach of Cunninghamia lanceolata breeding measures in Fujian province[J].China Forestry Science and Technology,2008,22(2):1-6.
[5] 何祯祥,施季森,王明庥,等.杉木杂种群体分子框架遗传连锁图初报[J].南京林业大学学报,2000,24(6):22-26.He Z X,Shi J S,Wang M X,et al.The construction of molecular linkage frame map in Chinese fir[J].Journal of Nanjing Forestry University,2000,24(6):22-26.
[6] 柳李旺,龚义勤,黄浩,等.新型分子标记——SRAP与TRAP及其应用[J].遗传,2004,26(5):777-781.Liu L W,Gong Y Q,Huang H,et al.Novel molecular marker systems—SRAP and TRAP and their application[J].Hereditas,2004,26(5):777-781.
[7] Li G,Quiros C F.Sequence-related amplified polymorphism(SRAP),a new marker system based on a simple PCR reaction:its application to mapping and gene tagging in Brassica[J].Theoretical and Applied Genetics,2001,103(2):455-461.
[8] 邱帅,丁信誉,席梦利,等.东方百合SRAP体系优化及引物筛选[J]. 南京林业大学学报:自然科学版,2013,37(3):6-10.Qiu S,Ding X Y,Xi M L,et al.Optimization of SRAP-PCR system and primers screening of Oriental lily hybrids[J].Journal of Nanjing Forestry University:Natural Sciences Edition,2013,37(3):6-10.
[9] 李建军,刘志坚,肖层林,等.SRAP技术在遗传的研究进展[J].现代生物医学进展,2007,7(5):783-786.Li J J,Liu Z J,Xiao C L,et al.Research progress on SRAP technique in genetics [J].Progress in Modern Biomedicine,2007,7(5); 783-786.
[10] Sun S J,Gao W,Lin S Q,et al. Analysis of genetic diversity in Ganoderma population with a novel molecular marker SRAP [J].Applied Microbiology and Biotechnology,2006,72(3):537-543.
[11] 徐操,赵宝华.SRAP分子标记的研究进展及其应用 [J].生命科学仪器,2009,7(4):24-27.Xu C,Zhao B H.The development and application of SRAP molecular markers[J].Life Science Instruments,2009,7(4):24-27.
[12] 孙佳琦,梁建国,石少川,等.SRAP标记在观赏植物遗传育种中的应用[J].分子植物育种,2010,8(3):577-588.Sun J Q,Liang J G,Shi S C,et al.Applications of SRAP marker in geneticsand breeding of ornamental plants[J].Molecular Plant Breeding,2010,8(3): 577-588.
[13] 吴莺莺,彭方仁,郝明灼,等.油茶SRAP-PCR反应体系的建立[J].南京林业大学学报:自然科学版,2011, 35(5):112-116.Wu Y Y,Peng F R,Hao M Z,et al.Establishment reaction system of the SRAP-PCR in Camellia oleifera [J].Journal of Nanjing Forestry University:Natural Sciences Edition,2011,35(5):112-116.
[14] 李慧芝,尹燕枰,张春庆,等.SRAP在葱栽培品种遗传多样性研究中的适用性分析[J].园艺学报,2007,34(4):929-934.Li H Z,Yin Y P,Zhang C Q,et al.Evaluation of application of SRAP on analysis of genetic diversity in cultivars of Allium fistulosum L.[J].Acta Horticulturae Sinica,2007, 34(4):929-934.
[15] 丁锦平,高国庆,周瑞阳,等.花生SRAP反应体系的建立与优化[J].安徽农业科学,2008,36(10):4009-4010.Ding J P,Gao G Q,Zhou R Y,et al.Establishment and optimization of SRAP reaction conditions for Arachis hypogaea L.[J].Journal of Anhui Agricultural Sciences,2008,36(10):4009-4010.
[16] 张大伟,马林,陈全家,等.棉花SRAP反应体系的建立与优化[J].新疆农业大学学报,2008,31(5):23-27.Zhang D W,Ma L,Chen Q J,et al.Establishment and optimization of reaction system of sequence-related amplified polymorphism of cotton[J].Journal of Xinjiang Agricultural University,2008,31(5):23-27.
[17] Elizabethá Claxton E.Synthesis of allenes by 2,3-sigmatropic rearrangement of prop-2-yn-1-yl oxonium ylides formed in rhodium(II)carboxylate catalyzed-reactions of diazo-compounds [J].J Chem Soc-Chem Commun,1990(1):46-48.
[18] 何正文,刘运生,陈立华,等.正交设计直观分析法优化PCR条件[J].湖南医科大学学报,1998,23(4): 76-77.He Z W,Liu Y S,Chen L H,et al.Optimization of PCR conditions direct analysis based on orthogonal design[J].Bulletin of Hunan Medical University,1988,23(4):76-77.
[19] 朱红兵,席凯强.SPSS 17.0中的正交试验设计与数据分析[J].首都体育学院学报,2013,25(3):283-288.Zhu H B,Xi K Q.The orthogonal experimental design in SPSS17.0 and data analysis[J].Journal of Capital University of Physical Education and Sports,2013,25(3):283-288.
[20] 盛文涛,周劲松,汤泳萍,等.芦笋SRAP反应体系优化及引物筛选[J].分子植物育种,2010,8(3):612-618.Sheng W T,Zhou J S,Tang Y P,et al.Optimization of SRAP reaction system and selection of primers for Asparagus officinalis L.[J].Molecular Plant Breeding,2010,8(3):612-618.
[21] 邹明宏,郭凌飞,曾辉,等.山竹子基因组DNA提取及SRAP反应体系优化[J].经济林研究,2009,27(3): 38-41.Zou M H,Guo L F,Zeng H,et al.Genome DNA extraction and SRAP reaction system optimization of Garcinia mangostana [J].Nonwood Forest Research,2009,27(3): 38-41.
[22] 井赵斌,俞靓,魏琳,等.本氏针茅SRAP-PCR反应体系的建立及引物筛选[J].草业科学,2012,29(2):219-228.Jing Z B,Yu J,Wei L,et al.Establishment of SRAP-PCR system and primers screening of Stipa bungeana[J].Pratacultural Science,2012,29(2):219-228.
[23] 姜艳,刘进平.胡椒SRAP反应体系的建立和优化[J]. 中国农学通报,2012,28(31):141-145.Jiang Y,Liu J P.Establishment and optimization of black pepper SRAP reaction system[J].Chinese Agricultural Science Bulletin,2012,28(31):141-145.
[24] Wang G,Pan J,Li X,et al.Construction of a cucumber genetic linkage map with SRAP markers and location of the genes for lateral branch traits [J].Science in China Series C:Life Sciences,2005,48(3):213-220.
[25] 姜树坤,马慧,刘君,等.利用SRAP标记分析玉米遗传多样性[J].分子植物育种,2007,5(3):412-416.Jiang S K,Ma H,Liu J,et al. Genetic diversity in maize inbred lines revealed by SRAPs marker [J]. Molecular Plant Breeding,2007,5(3):412-416.

PDF(2712803 KB)

Accesses

Citation

Detail

Sections

Recommended

The full text is translated into English by AI, aiming to facilitate reading and comprehension. The core content is subject to the explanation in Chinese.

www.nldxb.njfu.edu.cn

Edited ＆ Published by Editorial Department of Nanjing Forestry University(Natural Sciences Edition)
Address： No.159 Longpan Road,Nanjing 210037 Jiangsu,P.R.China
E-mail ：xuebao@njfu.edu.cn , xuebao@njfu.com.cn
Telephone +86-25-85428247,85427076
Distributed by China International Book Trading Corporation P.O. Box 399, Beijing ,P.R. China

〈

〉

Please choose a citation manager

Content to export