Establishment and Optimization of SRAP -PCR Reaction System in Radish

doi:10.7668/hbnxb.2009.S1.023

Abstract

Abstract: In this study,radish genomic DNA was used for the PCR template to optimize radish SRAP -PCR reaction system.A SRAP -PCR system with good stability was established.In the 20 μL reaction system ,DNA 50 ng ,Mg2+2.25mmol/L,dNTP 0.2 mmol/L,Taq 1 U,Primer 0.7 μmol/L.PCR procedure :initially denaturing at 94 ℃ for 5 min ;then94 ℃, 1 min,35 ℃, 1 min and 72 ℃, 1 min for the first five cycles,then the annealing temperature was raised to 50 ℃ for another 35 cycles;At last,72 ℃10min and 4 ℃for ever.The optimum SRAP -PCR reaction system is adaptive to different radish variety.It laid a good foundation for genetic diversity analysis,genetics map construction and molecular assisted breeding in radish.

Key words: Radish(RaphanussativusL.), SRAP-PCR, Reactionsystem

CLC Number:

S631.03

LIHong-shuang, QIUYang, LIXi-xiang. Establishment and Optimization of SRAP -PCR Reaction System in Radish[J]. ACTA AGRICULTURAE BOREALI-SINICA, doi: 10.7668/hbnxb.2009.S1.023.

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References

[1] Lic G F.Quiros.Sequence -related amplified polymorphism(SRAP),a new marker system base on a simple PCR reaction:its application to mapping and gene tagging in Brassica[J].Theor Appl Gene,2001,103 :455 -461.
[2] Riaz A,Potter D,Stephen M.Genotyping of peach and nectarinecultivars with SSR and SRAP molecular markers[J].JAmer Soc Hort Sci,2004,129:204-211.
[3] Ferriol M,Pico B,Nuez F.Genetic diversity of a 　germplasmcollection of Cucurbita pepo using SRAP and AFLP markers[J].Theor Appl Genet,2003,107:271 -282.
[4] Lin Z X,Zhang X L,Nie Y C,et al.Construction of a geneticlinkagemap for cotton based on SRAP[J].Chinese ScienceBulletin,2003,48(19):2063-2067.
[5] Li G,Gao M,Yang B,et al.Gene for gene alignment betweenthe Brassica and Arabidopsis genomes by direct transcriptomemapping [J].Theor Appl Genet,2003,107(1):168 -180.
[6] 潘俊松,王　刚.黄瓜SRAP 遗传连锁图谱的构建及始花节位的基因定位[J].自然科学进展,2005,15(2):167-172.
[7] 武志朴,杨文香,刘大群,等.小麦基因组SRAP 扩增体系的初步研究[J].河北农业大学学报,2005,28(3):66-69.
[8] 任　羽,王得元,张银东,等.辣椒SRAP -PCR 反应体系的建立与优化[J].分子植物育种,2004,2(5):689 -693.
[9] 林忠旭,张献龙,聂以春,等.棉花SRAP 遗传连锁图谱的构建[J].科学通报,2003,48(15):90 -93.
[10] 梁景霞,祁建民,吴为人,等.烟草DNA 的提取与SRAP反应体系的建立[J].中国烟草学报,2005,11(4):33-38.
[11] 赵新亮,马　强.玉米SRAP 反应体系的建立与优化[J].安徽农业科学,2006,34(15):3619-3620.
[12] 彭　飒,郭美丽,陈跃华,等.红花SRAP 扩增体系的建立和优化[J].第二军医大学学报,2006,27(5):544 -547.
[13] Liu L,Guo W,Zhu X,et al.Inheritance and fine mapping offertility -restoration for cytoplasmic male sterility in Gosypiumhirsutum L[J].Theor Appl Genet,2003,106(3):461-469.
[14] 王　佳,梁国华.正交设计优化黄瓜ISSR 体系[J].分子植物育种,2005,4(3):439-422.
[15] 余　艳,陈海山,葛学军.简单重复序列区间(ISSR)引物反应条件优化与筛选[J].热带亚热带植物学报,2003,11(1):15 -19.
[16] 继九如,黄智慧,倪红伟.正交设计优化兴兴草ISSRPCR反应体系研究[J].中国草地学报,2006,28(6):52-55

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