CDNA FROM RNA OF NAVICULA SALINICOLA NLA DIATOM GROWN INDOOR AND OUTDOOR FOR SQRT-PCR APPLICATIONS

• Main Author: Audria, Trisha
• Format: Theses
• Language: Indonesia
• Subjects: Kimia
• Online Access: https://digilib.itb.ac.id/gdl/view/71599
• Institution: Institut Teknologi Bandung

Navicula salinicola NLA is one of the diatoms that has the potential to be used as biodiesel feedstock. Biodiesel production of N. salinicola NLA is affected by light intensity, and this can be studied from its lipid metabolism pathway. However, so far information on lipid metabolism pathways in N. salinicola NLA grown under various light intensity conditions is unknown. One way to identify lipid metabolism in N. salinicola NLA is to identify the level of RNA transcripts via cDNA by reverse transcription polymerase chain reaction (RT-PCR). The aim of this study was to obtain cDNA from RNA of N. salinicola NLA grown indoors and outdoors for sqRT-PCR applications. Research methods included indoor and outdoor cultivation of N. salinicola NLA, isolation of total lipids, design of primers for internal gene control ofribulose-bisphosphate carboxylase (rbcL) and ?-actin (ACTB), isolation of genomic DNA of N. salinicola NLA, testing of primer compatibility control internal genes on genomic DNA, total RNA isolation, cDNA synthesis from RNA, and evaluation of control internal genes on cDNA from N. salinicola RNA. The results showed that N. salinicola NLA cells were shaped like a boat with dark brown culture color for indoor culture and light yellowish brown for outdoor culture. The initial density of N. salinicola NLA cells was 0.75 × 106 cells mL–1 , which increased to 4.1 and 2.2 ×106 cells mL –1 after 6 days of cultivation indoors and outdoors, respectively. This corresponds to a specific growth rate of 0,33 ± 0,02 and 0,24 ± 0,03 × 104 cells mL –1 day –1, doubling time of 2,11 ± 0,12 and 2,94 ± 0,47 days, biomass productivity of 0,53 ± 0,01 and 0,23 ± 0,01mg mL–1 day–1, and yield of 0,56 ± 0,08 and 0,41 ± 0,04 g L–1 on sixth day.. The yield of lipids produced from N. salinicola NLA cultures cultivated indoors was 41.12%, whereas those grown outdoors yielded 46.1%. Internal control genes of the N. salinicola NLA genome were discovered from a 500-bp segment on NAVsa;RbcL;1 and ACTB;1, demonstrating that the primer is suitable. In the exponential and stationary phases of both indoor and outdoor-grown N. salinicola NLA, total RNA was recovered, and the presence of 1400, 900, and 250 bp bands indicating 28S, 18S, and 5S RNA bands with a concentration range of 800-100 ng L –1 was identified. In the amplification of cDNA from total RNA using internal control genes primers, two 500 bp DNA fragments of NAVsa;RbcL;1 and NAVsa;ACTB;1 were effectively identified. ACTB consistently expressed in both indoor and outdoor-grown N. salinicola NLA