France S C, Kocher T D
Department of Zoology, University of New Hampshire, Durham 03824, USA.
Mol Mar Biol Biotechnol. 1996 Dec;5(4):304-13.
Marine invertebrate collections have historically been maintained in ethanol following fixation in formalin. These collections may represent rare or extinct species or populations, provide detailed time-series samples, or come from presently inaccessible or difficult-to-sample localities. We tested the viability of obtaining DNA sequence data from formalin-fixed, ethanol-preserved (FFEP) deep-sea crustaceans, and found that nucleotide sequences for mitochondrial 16S rRNA and COI genes can be recovered from FFEP collections of varying age, and that these sequences are unmodified compared with those derived from frozen specimens. These results were repeatable among multiple specimens and collections for several species. Our results indicate that in the absence of fresh or frozen tissues, archived FFEP specimens may prove a useful source of material for analysis of gene sequence data by polymerase chain reaction (PCR) and direct sequencing.
海洋无脊椎动物标本历来是在福尔马林固定后保存在乙醇中。这些标本可能代表稀有或灭绝的物种或种群，提供详细的时间序列样本，或者来自目前无法到达或难以采样的地点。我们测试了从福尔马林固定、乙醇保存（FFEP）的深海甲壳类动物中获取DNA序列数据的可行性，发现线粒体16S rRNA和COI基因的核苷酸序列可以从不同年代的FFEP标本中获得，并且与从冷冻标本中获得的序列相比，这些序列没有被修饰。这些结果在多个物种的多个标本和样本中是可重复的。我们的结果表明，在没有新鲜或冷冻组织的情况下，存档的FFEP标本可能是通过聚合酶链反应（PCR）和直接测序分析基因序列数据的有用材料来源。
