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为了保证玉米品种DNA指纹库构建的标准化,从建库标记、检测平台、试剂、样品、评估程序、数据整合、模式库、扩展库、随机盲测等九个方面进行了全面的规范:(1)通过对不同分子标记的比较,确定SSR标记作为建库标记;(2)通过对不同DNA指纹检测方法的比较,确定毛细管电泳结合多色荧光检测方法作为DNA指纹片段分析方法;(3)规范了DNA、引物、Taq酶等试剂质量,以保证数据的稳定性和可重复性;(4)规范了样品来源、样品类型及分析样品数量;(5)确定评估品种的数量及引物的取舍标准;(6)为解决不同来源DNA指纹数据的有效整合问题,提出了固定一套核心引物、提供标准品种、提供标准DNA、确定引物等位基因BIN、规定对异常带型数据处理方式、规定数据的编码方式等6个解决策略;(7)规范了构建模式库的材料名单、来源及构建方式;(8)规范了构建扩展库的材料来源及构建方式;(9)进行数据库数据随机盲测,以评估入库数据质量。以上规范将为不同单位合作开展大规模的DNA指纹库构建研究奠定基础。
In order to ensure the standardization of DNA fingerprinting in maize cultivars, a comprehensive specification was made from nine aspects including library marking, testing platform, reagents, samples, evaluation procedures, data integration, pattern library, extension library and random blind test. ) By comparing different molecular markers to determine SSR markers as a library marker; (2) by comparing different DNA fingerprinting methods to determine the capillary electrophoresis combined with multi-color fluorescence detection method as DNA fingerprinting method; (3) (4) regulate the sample source, sample type and analysis of the number of samples; (5) to determine the number of varieties and primers to assess the selection criteria ; (6) In order to solve the problem of effective integration of DNA fingerprinting data from different sources, a set of core primers, a set of standard primers, a standard DNA set, a BIN set of primer alleles, a set of abnormal band data processing methods, (7) standardize the list of materials to build a model library, sources and methods of construction; (8) standardize the source and structure of building an extension of the library Manner; (9) a random database blind test data to evaluate the quality of the data storage. The above specifications will lay the foundation for the large-scale research on the construction of DNA fingerprinting libraries by different units.