摘要： 应用RAPD与dpRAPD方法鉴定萝卜-甘蓝型油菜中萝卜基因组,筛选了10条随机引物.结果表明,平均每条引物(组合)能产生的萝卜基因组特异标记数dpRAPD高于RAPD,分别为1.69和1.33;在dpRAPD扩增产物中有77.6%谱带清晰易辨,略高于RAPD(75.%).两者所检测到的萝卜基因组标记大部分为各自特异的扩增产物.由于结合了荧光标记引物,dpRAPD反应产物可在Genetic Analyzer上分离检测,因此能检测到100bp以下的小片段DNA.
关键词:
RAPD,
dpRAPD,
聚丙烯酰胺凝胶电泳,
银染
Abstract: Normally, a single 10-mer oligonucleotide primer is used in a RAPD amplification. However, if pairs of two primers are used, theoretically more fragments should be amplified. In this experiment, we used 140 decamer primers to screen molecular markers specific for individual radish chromosomes in Raphanobrassica, both individually and in pairs (here named dpRAPD) with three fluorescence-labelled primers: OPD11FAM, OPG19Tet and OPH15Hex. On the average, each single primer generated 1.3 chromosome-specific markers in RAPD, while each pairwise combination of primers generated 1.7 markers in dpRAPD. The percentage of excellent and good bands in dpRAPD (78% ) was higher than for RAPD (75%). It was found that the majority of dpRAPD bands are different from original RAPD bands. Fragments with length under 100 bp which are difficult to be identified by polyacrylamide or agarose gel electrophoresis are detectable using Genetic Analyzer. It was shown that dpRAPD is a useful complement to classical RAPD analysis.
Key words:
RAPD,
dp RAPD,
Polyacrylamide gel electrophoresis,
Silver staining
中图分类号:
丁云花, HolgerBudahn, 简元才, HerhertPeterka. 萝卜-甘蓝型油菜中萝卜基因组的RAPD与dpRAPD分析效率研究[J]. 华北农学报, 2004, 19(2): 20-23. doi: 10.3321/j.issn:1000-7091.2004.02.006.
DING Yun-hua, Holger Budahn, JIAN Yuan-cai, Herbert Peterka. Efficiency of RAPD and Double Primer (dp)RAPD for Detection of Radish-genomic Components in Raphanobrassica[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2004, 19(2): 20-23. doi: 10.3321/j.issn:1000-7091.2004.02.006.