枣ISSR反应体系的建立及其指纹图谱构建

Establishment of inter-simple sequence repeat(ISSR) reaction system and construction of fingerprint in Ziziphus jujuba genus

对影响枣ISSR反应的相关因子进行了研究。优化的反应体系包含:2.5μL 10×buffer,50 ng模板DNA,2.5 mmol.L-1 Mg2+,0.3 mmol.L-1 dNTPs,0.3μmol.L-1引物,2 U Taq DNA聚合酶,反应体积为25μL;优化的PCR扩增程序为:94℃2 min;94℃l min,退火1 min,72℃2 min,循环35次;72℃延伸10 min;退火温度根据引物温度设定。该ISSR体系被用于17个枣栽培品种遗传图谱分析,结果表明所构建的遗传图谱能较好地反应供试品种的遗传多样性。

Several main factors influencing inter-simple sequence repeat（ISSR） reaction were analyzed in order to establish ISSR molecular marker system in Zizyphus jujube genus.The optimal system was in a reaction volume of 25 μL including： 2.5 ？L 10×buffer,50 ng template DNA,2.5 mmol·L-1 Mg2＋,0.3 mmol·L-1 dNTPs,0.3 μmol·L-1 primer,2 U TaqDNA polymerase.The optimal amplification program was with the following cycling parameters： 94°C for 2 min;35 cycles of 94°C for 1 min;specified annealing temperature for 1 min,72°C for 2 min,final extension at 72°c for 10 min.The optimal amplification program was used in the present study to detect the genetic diversity in 17 cultivars of Jujube.