Premium
Exploiting formalin‐preserved fish specimens for resources of DNA barcoding
Author(s) -
ZHANG J.
Publication year - 2010
Publication title -
molecular ecology resources
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.96
H-Index - 136
eISSN - 1755-0998
pISSN - 1755-098X
DOI - 10.1111/j.1755-0998.2010.2838.x
Subject(s) - biology , dna barcoding , barcode , genomic dna , dna , polymerase chain reaction , gene , multiple displacement amplification , computational biology , dna extraction , microbiology and biotechnology , genetics , evolutionary biology , computer science , operating system
With the development of the DNA barcoding project, a large number of specimens are required to establish the library of reference barcode. Formalin‐fixed samples from museums provide a potential resource for it. However, recovery of DNA and amplification of the target gene from formalin‐fixed samples are challenging. In this study, a hot alkali pre‐treatment accompanied by the use of cetyltrimethylammonium bromide ( CTAB) method was employed for DNA recovery from formalin‐preserved samples, with the purpose of pursuing the optimal condition for high quantity and quality of DNA and minimizing PCR inhibition. Meanwhile, a semi‐nested PCR‐based method was developed to enhance the efficacy of amplification. This advanced protocol was demonstrated to be reliable and effective. Even for 23‐year‐old samples, genomic DNA could be extracted, and COI gene was correctly sequenced.