The paper aimed to determine the true identity of a presumed or commonly believed Sinarapan fish and compare their phylogenetic relationships by using DNA barcoding. The fish samples were collected by researchers from...The paper aimed to determine the true identity of a presumed or commonly believed Sinarapan fish and compare their phylogenetic relationships by using DNA barcoding. The fish samples were collected by researchers from a fisheries research and development institution in April 2015 from four different lakes of Bicol Region, i.e., Lakes Buhi, Danao, Manapao and Bato. DNA was extracted using cetyl trimethyl ammonium bromide （CTAB） extraction buffer. The polymerase chain reaction （PCR） profile for the reaction was 94 ℃ for 10 min, followed by 35 cycles of 1 min at 94℃, 1 min at 48℃ and 1.5 min at 72℃, and a final extension of 10 min at 72℃. The CO 1 region with approximately 650 bp was amplified because of its capability to differentiate taxa. Sequencing was done by Macrogen while phylogenetic analysis was produced using a Molecular Evolutionary Genetics Analysis （MEGA） software version 6.0. The findings showed that CO1 can be used as a DNA marker in molecular identification of the fish samples. Samples from two of the four lakes were successfully sequenced. From basic local alignment search tool （BLAST） results, the maximum parsimony （MP） and neighbor-joining （N J） trees revealed that samples from Lakes Buhi and Bato are not species of Sinarapan but rather species of Leiopotherapon plumbeus and Rhinogobius giurinus, respectively. Furthermore, DNA barcoding is very useful in proving the true identity of unknown samples.展开更多
文摘The paper aimed to determine the true identity of a presumed or commonly believed Sinarapan fish and compare their phylogenetic relationships by using DNA barcoding. The fish samples were collected by researchers from a fisheries research and development institution in April 2015 from four different lakes of Bicol Region, i.e., Lakes Buhi, Danao, Manapao and Bato. DNA was extracted using cetyl trimethyl ammonium bromide （CTAB） extraction buffer. The polymerase chain reaction （PCR） profile for the reaction was 94 ℃ for 10 min, followed by 35 cycles of 1 min at 94℃, 1 min at 48℃ and 1.5 min at 72℃, and a final extension of 10 min at 72℃. The CO 1 region with approximately 650 bp was amplified because of its capability to differentiate taxa. Sequencing was done by Macrogen while phylogenetic analysis was produced using a Molecular Evolutionary Genetics Analysis （MEGA） software version 6.0. The findings showed that CO1 can be used as a DNA marker in molecular identification of the fish samples. Samples from two of the four lakes were successfully sequenced. From basic local alignment search tool （BLAST） results, the maximum parsimony （MP） and neighbor-joining （N J） trees revealed that samples from Lakes Buhi and Bato are not species of Sinarapan but rather species of Leiopotherapon plumbeus and Rhinogobius giurinus, respectively. Furthermore, DNA barcoding is very useful in proving the true identity of unknown samples.
