CIRAD, UMR AGAP, F-34398 Montpellier, France.
Am J Bot. 2012 Jun;99(6):e245-50. doi: 10.3732/ajb.1100548. Epub 2012 May 29.
Discrepancies in terms of genotyping data are frequently observed when comparing simple sequence repeat (SSR) data sets across genotyping technologies and laboratories. This technical concern introduces biases that hamper any synthetic studies or comparison of genetic diversity between collections. To prevent this for Sorghum bicolor, we developed a control kit of 48 SSR markers.
One hundred seventeen markers were selected along the genome to provide coverage across the length of all 10 sorghum linkage groups. They were tested for polymorphism and reproducibility across two laboratories (Centre de Cooperation Internationale en Recherche Agronomique pour le Developpement [CIRAD], France, and International Crops Research Institute for the Semi-Arid Tropics [ICRISAT], India) using two commonly used genotyping technologies (polyacrylamide gel-based technology with LI-COR sequencing machines and capillary systems with ABI sequencing apparatus) with DNA samples from a diverse set of 48 S. bicolor accessions.
A kit for diversity analysis (http://sat.cirad.fr/sat/sorghum_SSR_kit/) was developed. It contains information on 48 technically robust sorghum microsatellite markers and 10 DNA controls. It can further be used to calibrate sorghum SSR genotyping data acquired with different technologies and compare those to genetic diversity references.
当比较不同基因分型技术和实验室的简单重复序列(SSR)数据集时,经常会观察到基因分型数据的差异。这种技术上的差异会引入偏差,从而阻碍任何综合研究或不同收集品系之间遗传多样性的比较。为了防止这一点在高粱中发生,我们开发了一个 48 个 SSR 标记的对照试剂盒。
沿着基因组选择了 117 个标记,以提供对所有 10 个高粱连锁群全长的覆盖。它们使用两种常用的基因分型技术(基于聚丙烯酰胺凝胶的技术与 LI-COR 测序仪和带有 ABI 测序仪的毛细管系统)在两个实验室(法国国际农业研究磋商组织 [CIRAD]和国际半干旱热带作物研究所 [ICRISAT])进行了多态性和可重复性测试,使用来自多样化的 48 个高粱品种的 DNA 样本。
开发了一个用于多样性分析的试剂盒(http://sat.cirad.fr/sat/sorghum_SSR_kit/)。它包含了 48 个技术上可靠的高粱微卫星标记和 10 个 DNA 对照的信息。它还可以用于校准不同技术获得的高粱 SSR 基因分型数据,并将其与遗传多样性参考进行比较。