在龙葵杂交群体中检测到随机扩增多态性DNA标记

K. Hosaka, R. Hanneman
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引用次数: 23

摘要

通过调节扩增过程的温度分布和模板DNA浓度(反应体积为1 ng/μl),对两个马铃薯品种(沙香茄(Solanum chacoense)和普雷加(S. phureja)的一个F1群体及其衍生的F2群体进行随机扩增,获得了可重复的随机扩增多态性DNA (RAPD)图谱。虽然随着引物GC含量的增加,扩增产物的数量和两种之间可检测到的差异增加,但建议在F2群体中50%或60% GC可以最大化可评分的rapd。利用82条引物,在亲本间共检测到589个rapd,其中70%(409个rapd)发生在F1克隆中。由于复杂的RAPD模式,在F2群体中可靠评分的RAPD数量显著降低。结果,在亲本中检测到的rapd中,有22%(129个)被作为遗传标记,平均每个引物有1.57个rapd,可用于构建该特定群体的遗传图谱。
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Random amplified polymorphic DNA markers detected in a segregating hybrid population of Solanum chacoense × S. phureja
Reproducible random amplified polymorphic DNA (RAPD) patterns were obtained in two potato species (Solanum chacoense and S. phureja), one of their F1's, and its derived F2 population, by adjusting temperature profiles of the amplification process and template DNA concentrations (1 ng/μl reaction vol.). Although the number of amplified products and detectable differences between the two species increased with increasing GC content of the primer, 50% or 60% GC is recommended for maximizing scorable RAPDs in the F2 population. Using 82 primers, 589 RAPDs were detected between the parents, and 70% of them (409 RAPDs) in the F1 clone. The number of RAPDs reliably scored for their segregations in the F2 population was significantly lowered because of complicated RAPD patterns. Consequently, 22% of the RAPDs detected in the parents (129 RAPDs), an average of 1.57 RAPDs per primer, were obtained as genetic markers, which can be used for the construction of a genetic map for this particular population.
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