Moss Landing Marine Laboratories, 8272 Moss Landing Road, Moss Landing, CA 95309, USA.
Mol Ecol Resour. 2013 Sep;13(5):851-61. doi: 10.1111/1755-0998.12138. Epub 2013 Jul 13.
DNA barcoding is a powerful tool for species detection, identification and discovery. Metazoan DNA barcoding is primarily based upon a specific region of the cytochrome c oxidase subunit I gene that is PCR amplified by primers HCO2198 and LCO1490 ('Folmer primers') designed by Folmer et al. (Molecular Marine Biology and Biotechnology, 3, 1994, 294). Analysis of sequences published since 1994 has revealed mismatches in the Folmer primers to many metazoans. These sequences also show that an extremely high level of degeneracy would be necessary in updated Folmer primers to maintain broad taxonomic utility. In primers jgHCO2198 and jgLCO1490, we replaced most fully degenerated sites with inosine nucleotides that complement all four natural nucleotides and modified other sites to better match major marine invertebrate groups. The modified primers were used to amplify and sequence cytochrome c oxidase subunit I from 9105 specimens from Moorea, French Polynesia and San Francisco Bay, California, USA representing 23 phyla, 42 classes and 121 orders. The new primers, jgHCO2198 and jgLCO1490, are well suited for routine DNA barcoding, all-taxon surveys and metazoan metagenomics.
DNA 条形码是一种用于物种检测、鉴定和发现的强大工具。后生动物 DNA 条形码主要基于细胞色素 c 氧化酶亚基 I 基因的特定区域，该区域通过 Folmer 等人设计的引物 HCO2198 和 LCO1490（“Folmer 引物”）进行 PCR 扩增。（分子海洋生物学和生物技术，3，1994，294）。自 1994 年以来发表的序列分析表明，许多后生动物的 Folmer 引物存在不匹配。这些序列还表明，更新的 Folmer 引物需要极高水平的简并性才能保持广泛的分类学用途。在引物 jgHCO2198 和 jgLCO1490 中，我们用肌苷核苷酸替换了大多数完全简并的位点，肌苷核苷酸与所有四个天然核苷酸互补，并修饰了其他位点以更好地匹配主要的海洋无脊椎动物群。用修改后的引物从来自法属波利尼西亚的莫雷阿岛和美国加利福尼亚州旧金山湾的 9105 个标本中扩增和测序细胞色素 c 氧化酶亚基 I，这些标本代表 23 个门、42 个纲和 121 个目。新的引物 jgHCO2198 和 jgLCO1490 非常适合常规 DNA 条形码、全分类调查和后生动物宏基因组学。
