Five-color-based high-information-content fingerprinting of bacterial artificial chromosome clones using type IIS restriction endonucleases
Five-color-based high-information-content fingerprinting of bacterial artificial chromosome clones using type IIS restriction endonucleases
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DOI：
10.1006/geno.2001.6547
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发表时间：
2001-06-01
期刊：
影响因子：
4.4
通讯作者：
Shizuya, H
中科院分区：
文献类型：
--
作者：
Ding, Y;Johnson, MD;Shizuya, H
我们开发了一种高信息含量的细菌人工染色体(BAC)克隆的指纹分析系统，该系统使用IIS型限制性内切酶HGaI和II型限制性内切酶RsaI。在所描述的方法中，用HGaI产生的未知的五个碱基突出物被修饰的荧光双脱氧终止子部分或全部测序。使用用第五种染料标记的泳道内大小标准，片段的特征是其末端一到五个碱基的大小和序列。与该方法相关的增强的信息内容显著提高了检测BAC克隆之间共享片段的准确性和效率。我们已经将这种方法得到的数据与预测的10个全序列BAC的HICF模式进行了比较。我们进一步将HICF应用于555个BAC克隆，以组装跨越人类16号染色体16p11.2至16p13.1的重叠群。(C)2001年学术出版社。
We have developed a high-information-content fingerprinting (HICF) system for bacterial artificial chromosome (BAC) clones using a Type IIS restriction endonuclease, HgaI, paired with a Type II restriction endonuclease, RsaI. In the method described, unknown five-base overhangs generated with HgaI are partially or fully sequenced by modified fluorescent dideoxy terminators. Using an in-lane size standard labeled with a fifth dye, fragments are characterized by both the size and the sequence of its terminal one to five bases. The enhanced information content associated with this approach significantly increases the accuracy and efficiency of detecting shared fragments among BAC clones. We have compared data obtained from this method to predicted HICF patterns of 10 fully sequenced BACs. We have further applied HICF to 555 BAC clones to assemble contigs spanning 16p11.2 to 16p13.1 of human chromosome 16. (C) 2001 Academic Press.