Research Article
The Use of Degenerate Primer to Isolation and Designing Housekeeping Gene of Eel Fish (Anguilla bicolor)
@INPROCEEDINGS{10.4108/eai.12-10-2019.2296368, author={Diah Kusumawaty and Tina Yulianti and Hernawati Hernawati}, title={The Use of Degenerate Primer to Isolation and Designing Housekeeping Gene of Eel Fish (Anguilla bicolor)}, proceedings={Proceedings of the 7th Mathematics, Science, and Computer Science Education International Seminar, MSCEIS 2019, 12 October 2019, Bandung, West Java, Indonesia}, publisher={EAI}, proceedings_a={MSCEIS}, year={2020}, month={7}, keywords={anguilla bicolor housekeeping gene degenerate primer nested pcr specific primer}, doi={10.4108/eai.12-10-2019.2296368} }- Diah Kusumawaty
Tina Yulianti
Hernawati Hernawati
Year: 2020
The Use of Degenerate Primer to Isolation and Designing Housekeeping Gene of Eel Fish (Anguilla bicolor)
MSCEIS
EAI
DOI: 10.4108/eai.12-10-2019.2296368
Abstract
This study aims to sequence the isolation results of the housekeeping gene in Anguilla bicolor to get specific sequences by designing degenerate primer on 18s ribosomal RNA (18S rRNA), Beta Actin (ACTB), Elongation Factor 1-Alpha (ef1a) and Glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH). The degenerate primer consists of two primer sets (outer and inner) that were designed by the Primaclade method. The result of housekeeping gene amplification was then sequenced directly to determine the nucleotide sequences of the gene. The results of BLAST analysis showed that the three of four genes sequences were corresponding to the target gene, which are ribosome RNA 18s, (ACTB) and Ef1a. The general grouping of dendrograms showed that the three gene sequences were in groups with a gene target. The conclusion is the discovery of three specific sequences in three housekeeping gene targets which can be used as a source for designing specific primers in subsequent studies.