Improved discrimination of self-incompatibility S-RNase alleles in cherry and high throughput genotyping by automated sizing of first intron polymerase chain reaction products
Improved discrimination of self-incompatibility S-RNase alleles in cherry and high throughput genotyping by automated sizing of first intron polymerase chain reaction products
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DOI：
10.1111/j.1439-0523.2006.01205.x
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发表时间：
2006-06-01
期刊：
影响因子：
2
通讯作者：
Tobutt, K. R.
中科院分区：
文献类型：
--
作者：
Sonneveld, T.;Robbins, T. P.;Tobutt, K. R.
报道了一种新的聚合酶链反应（PCR）方法，以确定和确认樱桃自交不亲和（S）基因型。该方法包括用一对新的共有引物进行PCR扩增，该引物紧邻樱桃S-RNases的第一个内含子，其中一个是荧光标记的。荧光扩增产物范围从234到c。460 bp，并且可以在自动测序仪上精确地确定大小。甜樱桃中报道的13个S等位基因可以区分，除了S-2和S-7，它们具有完全相同大小的扩增产物。S-13，这也是扩增，给出了一个微卫星样的痕迹，显示微小的等位基因内长度的变化。该方法给出了快速、准确的结果，对于野生和栽培樱桃的中/高通量基因分型特别有用。
A novel polymerase chain reaction (PCR) approach to determine and confirm the self-incompatibility (S) genotype of cherries is reported. The method involves PCR amplification with a new pair of consensus primers that immediately flank the first intron of cherry S-RNases, one of which is fluorescently labelled. Fluorescent amplification products range from 234 to c. 460 bp and can be sized accurately on an automated sequencer. Thirteen S alleles reported in sweet cherry can be distinguished, except for S-2 and S-7, which have an amplification product of exactly the same size. S-13, which is also amplified, gives a microsatellite-like trace which shows minor intra-allelic length variation. This method gives fast and accurate results and should be especially useful for medium/high-throughput genotyping of wild and cultivated cherries.