Rapid isolation of CA microsatellites from the tilapia genome
Rapid isolation of CA microsatellites from the tilapia genome
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DOI：
10.1046/j.1365-2052.2002.00817.x
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发表时间：
2002-04-01
期刊：
影响因子：
2.4
通讯作者：
Kocher, TD
中科院分区：
文献类型：
--
作者：
Carleton, KL;Streelman, JT;Kocher, TD
我们已经开发了（CA），微卫星标记的慈鲷鱼。尼罗罗非鱼杂交捕获方法的变异。所得基因组文库高度富集重复DNA，其中96%的克隆含有CA重复序列。重复次数从4到45不等，平均为19。三分之二的测序克隆具有12个或更多的重复序列和足够的侧翼序列来设计引物。在O. niloticus x O.金黄色。近90%的标记扩增在这个交叉和74%的这些是信息。这项工作证明了在富集步骤之前和之后最小化聚合酶链反应（PCR）扩增循环的数量以减少PCR重组和嵌合克隆的产生的重要性。
We have developed (CA), microsatellite markers for the cichlid fish. Oreochromis niloticus using a variation of the hybrid capture method. The resulting genomic library was highly enriched in repetitive DNA with 96%, of clones containing CA repeats. The number of repeats ranged from four to 45 with an average of 19. Two-thirds of the sequenced clones had 12 or more repeats and sufficient flanking sequence to design primers. The resulting markers were tested in an F, cross of O. niloticus x O. aureus. Nearly 90% of the markers amplified in this cross and 74% of these were informative. This work demonstrates the importance of minimizing the number of polymerase chain reaction (PCR) amplification cycles before and after the enrichment steps to reduce PCR recombination and the generation of chimaeric clones.