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Jurnal Bioteknologi & Biosains Indonesia (JBBI)
Published by Badan Pengkajian dan Penerapan Teknologi
ISSN : 24422606     EISSN : 2548611X     DOI : -
Core Subject : Science, Agriculture,
Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Immunology & microbiology
JBBI, Indonesian Journal of Biotechnology & Bioscience, is published twice annually and provide scientific publication medium for researchers, engineers, practitioners, academicians, and observers in the field related to biotechnology and bioscience. This journal accepts original papers, review articles, case studies, and short communications. The articles published are peer-reviewed by no less than two referees and cover various biotechnology subjects related to the field of agriculture, industry, health, environment, as well as life sciences in general. Initiated at the then Biotech Centre, the journal is published by the Laboratory for Biotechnology, the Agency for the Assessment and Application of Technology, BPPT.
Arjuna Subject : -
Articles 542 Documents
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MIKROPROPAGASI TANAMAN TALAS BENENG (Xanthosoma undipes K. Koch) DENGAN PERLAKUAN BENZIL AMINOPURIN, TIAMIN, DAN ADENIN Sari, Laela; Wulansari, Aida; Noorrohmah, Siti; Ermayanti, Tri Muji
Jurnal Bioteknologi & Biosains Indonesia (JBBI) Vol 6, No 1 (2019): June 2019
Publisher : Badan Pengkajian dan Penerapan Teknologi (BPPT)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.29122/jbbi.v6i1.3216

Abstract

Micropropagation of Beneng Taro (Xanthosoma undipes K. Koch) with Benzyl Amino Purine, Thiamine, and Adenine TreatmentABSTRACTConventional production of Beneng taro seeds (Xanthosoma undipes K. Koch) is constrained by the limited number of tubers, thus an alternative solution is needed such as in vitro propagation. This study was aimed to obtain a micropropagation technique of Beneng taro on MS media with BAP, thiamine, and adenine treatment, and to determine its growth at the acclimatization stage. This research consisted of shoot multiplication and acclimatization. Shoot propagation was carried out on MS media with 8 treatments, namely ½MS and MS without addition of growth promoting substance, and MS with 1, 2 and 3 mg×L-1 BAP, with or without addition of 1 mg×L-1 thiamine and 2 mg×L-1 adenine. Each treatment was replicated four times, each consisting of four shoots. Growth observation was made from 1st to 5th week on petiole length, and number of shoots, leaves and roots. Acclimatization was carried out on soil media, compost, and husks in a ratio of 1: 1: 1. The results showed that the best media for shoot multiplication was MS + 1 mg×L-1 BAP + 1 mg×L-1 thiamine + 2 mg×L-1 adenine with an average of 3.5 shoots, while the best medium for the petiole length was ½MS with an average value of 6.97 cm. The results of acclimatization showed that 100% planlets survived, and plantlets grown on MS media + 3 mg×L-1 BAP had the highest number of shoots with an average of 4.2.Keywords: adenine, Beneng taro, benzil amino purine (BAP), micropropagation, thiamineABSTRAKPenyediaan bibit talas Beneng (Xanthosoma undipes K. Koch) secara konvensional terkendala terbatasnya jumlah umbi, sehingga perlu solusi alternatif, diantaranya melalui perbanyakan in vitro. Tujuan penelitian ini adalah untuk mendapatkan teknik mikropropagasi talas beneng pada media MS dengan perlakuan BAP, tiamin, adenin, dan untuk mengetahui pertumbuhannya pada tahap aklimatisasi. Penelitian ini meliputi perbanyakan tunas dan aklimatisasi. Perbanyakan tunas menggunakan media MS dengan 8 perlakuan yaitu ½MS dan MS tanpa penambahan zat pengatur tumbuh (ZPT), serta MS dengan 1, 2 dan 3 mg×L-1 BAP dengan atau tanpa penambahkan 1 mg×L-1 tiamin dan 2 mg×L-1 adenin. Setiap perlakuan mempunyai empat ulangan, setiap ulangan terdiri atas empat tunas. Pertumbuhan diamati mulai minggu ke-1 hingga ke-5 terhadap panjang petiol serta jumlah anakan, daun dan akar. Aklimatisasi dilakukan pada media tanah, kompos dan sekam dengan perbandingan 1:1:1. Hasil menunjukkan bahwa media terbaik perbanyakan tunas adalah MS + 1 mg×L-1 BAP + 1 mg×L-1 tiamin + 2 mg×L-1 adenin dengan rata-rata 3,5 tunas, sedangkan media terbaik untuk panjang tangkai daun adalah ½MS dengan nilai rata-rata 6,97 cm. Hasil aklimatisasi menunjukkan bahwa 100% planlet hidup dan planlet yang ditumbuhkan pada media MS + 3 mg×L-1 BAP mempunyai jumlah anakan terbanyak dengan rata-rata 4,2.Kata Kunci: adenine, benzil amino purin (BAP), mikropropagasi, talas Beneng, tiamine
TRANSFORMASI GENETIK DAN EKSPRESI MUTAN SUCROSE PHOSPHATE SYNTHASE PADA TANAMAN TOMAT Fibriani, Suwinda; Agustien, Inyana Dwi; Sawitri, Widhi Dyah; Sugiharto, Bambang
Jurnal Bioteknologi & Biosains Indonesia (JBBI) Vol 6, No 1 (2019): June 2019
Publisher : Badan Pengkajian dan Penerapan Teknologi (BPPT)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (699.256 KB) | DOI: 10.29122/jbbi.v6i1.3341

Abstract

Genetic Transformation and Expression of Sucrose Phosphate Synthase Mutant in Tomato Plant ABSTRACTSucrose phosphate synthase (SPS) is a key enzyme responsible for sucrose biosynthesis. In its regulation, SPS activity is modulated by an allosteric effector glucose-6-phosphate (G6P) suggested to have an ability to bind SPS N-terminus domain. To understand the role of N-terminus in regulating SPS, the SPS gene was mutated with the deletion of N-terminus domain (∆N-SPS). The ∆N-SPS gen was transformed into tomato plants with 5% transformation efficiency. Three transgenic tomato plant 4.20, 5.5.1, and 5.10 were obtained and confirmed by PCR analysis. Transgenic tomato expression was characterized by enzymatic analysis. Result showed that the G6P allosteric regulation in transgenic ∆N-SPS had lost and the SPS activity increased by 2-fold compared to non-transgenic plant. This showed that N-terminus domain-deleted SPS could be actively expressed in plant. Keywords: enzyme, genetic transformation, N-terminus domain deletion, sucrose phosphate synthase, tomato ABSTRAKSucrose phosphate synthase (SPS) merupakan enzim kunci yang bertanggung jawab dalam sintesis sukrosa. Dalam regulasinya, aktifitas SPS dipengaruhi oleh alosterik efektor glukosa-6-fosfat (G6P) yang diduga dapat berikatan pada domain N-terminus SPS. Untuk mengetahui peran N-terminus pada regulasi SPS, dilakukan mutasi SPS dengan penghilangan domain N-terminus (∆N-SPS). Gen ∆N-SPS diinsersi pada tanaman tomat melalui transformasi genetik dengan efisiensi transformasi 5%. Tiga tanaman transgenik tomat (event4.20; 5.5.1; dan 5.10) didapatkan dan positif terkonfirmasi melalui analisis PCR. Ekspresi mutan dikarakterisasi melalui analisis enzimatik. Hasil menunjukkan bahwa tanaman tomat transgenik ∆N-SPS tidak dipengaruhi regulasi alosterik G6P dan aktifitas SPS 2 kali lipat lebih tinggi daripada tanaman bukan transgenik. Ini menunjukkan bahwa SPS dengan delesi domain N-terminus dapat terekspresi aktif pada tanaman.  Kata Kunci: delesi domain N-terminus, enzim, sucrose phosphate synthase, tomat, transformasi genetik 
TELAAH METODE DIAGNOSIS CEPAT DAN PENGOBATAN INFEKSI Salmonella typhi Hardianto, Dudi
Jurnal Bioteknologi & Biosains Indonesia (JBBI) Vol 6, No 1 (2019): June 2019
Publisher : Badan Pengkajian dan Penerapan Teknologi (BPPT)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (542.569 KB) | DOI: 10.29122/jbbi.v6i1.2935

Abstract

Review on Rapid Diagnosis Method and Treatment of Salmonella typhi Infection ABSTRACTSalmonella is a genus of gram-negative bacilli which are pathogenic for human. Recently over 2,500 serotypes of Salmonella have been reported. Of these, the most common serotype causing typhoid fever which is acute infectious disease in small intestine due to S. typhi entering the body through contaminated food or drink. S. typhi infection remains a major public health concern worldwide because of the subsequent economic burden for the cost of surveillance, prevention, and treatment. In Indonesia, typhoid fever is an endemic disease that threatens public health and becomes a complex problem because it increases career cases and drug resistance, so its diagnosis is needed. Although there is already a diagnosis method of typhoid fever conventionally, a fast, easy and reliable diagnosis method is needed to diagnose typhoid fever by medical personnel in endemic countries. Typhoid fever is treated by antibiotics and prevention efforts are carried out through vaccination.Keywords: antibiotics, pathogen, rapid detection, Salmonella typhi, typhoid fever ABSTRAKSalmonella adalah bakteri basil gram negatif yang bersifat patogen terhadap manusia dan saat ini telah dilaporkan lebih dari 2.500 serotipe. Salah satu serotype Salmonella diketahui menyebabkan penyakit demam tifoid yaitu infeksi akut pada usus halus akibat S. typhi yang masuk ke dalam tubuh melalui makanan dan minuman yang tercemar. Infeksi S. typhi menjadi masalah utama dalam kesehatan masyarakat di seluruh dunia karena bebani ekonomi yang ditimbulkannya untuk biaya pengawasan, pencegahan, dan pengobatan. Di Indonesia, demam tifoid merupakan penyakit endemis yang mengancam kesehatan masyarakat dan menjadi masalah kompleks karena demam tifoid meningkatkan kasus-kasus karier dan resistensi obat sehingga diperlukan diagnosisnya. Meskipun sudah ada diagnosis demam tifoid secara konvensional, tetapi diperlukan metode diagnosis yang cepat, mudah dan andal untuk mendeteksi demam tifoid oleh tenaga medis yang bekerja di negara-negara endemik. Demam tifoid diobati dengan pemberian antibiotika dan dilakukan upaya pencegahan melalui vaksinasi.Kata Kunci: antibiotika, demam tifoid, deteksi cepat, patogen, Salmonella typhi
PRODUKSI LIPASE DARI ISOLAT KAPANG HASIL MUTASI UNTUK TRANSESTERIFIKASI Nabilasani, Galih Cendana; Siswodarsono, Trismilah; Suhendar, Dadang; Mubarik, Nisa Rachmania
Jurnal Bioteknologi & Biosains Indonesia (JBBI) Vol 6, No 1 (2019): June 2019
Publisher : Badan Pengkajian dan Penerapan Teknologi (BPPT)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (280.35 KB) | DOI: 10.29122/jbbi.v6i1.3047

Abstract

Lipase Production by Mutant Fungal Isolates for Transesterification ABSTRACTLipase is used amongst others in biodiesel production, namely in the transesterification reaction. Kernel B (KB) was a fungus isolated from the waste of palm kernel and seed. The fungus produced lipase that catalysed the transesterification reaction with a lower activity compared to that of AK Amano commercial lipase. The purpose of this study was to obtain mutant fungi with higher transesterification activities than the wild type (KB). The mutation process was carried out using ultraviolet (UV) light, ethyl methane sulfonate (EMS), and N-methyl-N’-nitro-N-nitrosoguanidine (NMNG) on KB fungus. The mutations using UV light produced 11 isolates, of which isolate m4.1KB1 produced a higher transesterification activity (0.172 U·mg-1) compared to the wild type. Mutant m5.7KB, which was generated from mutant m4.1KB1 treated using EMS, had its transesterification activity decreased to only 0.051 U·mg-1. Mutant m6.0,3KB2, which was resulted through NMNG treatment, experienced an increase in transesterification activity which was 91.2% higher than that of KB.Keywords: ethyl methane sulfonate, lipase, mutant fungi, N-methyl-N’-nitro-N-nitrosoguanidine, ultraviolet ABSTRAKLipase dimanfaatkan salah satunya dalam produksi biodiesel, yaitu dalam reaksi transesterifikasi. Kernel B (KB) merupakan kapang yang diisolasi dari limbah inti dan biji kelapa sawit, yang menghasilkan lipase sebagai katalis dalam reaksi transesterifikasi. Namun aktivitas transesterifikasi yang dihasilkan oleh lipase dari KB lebih rendah dibandingkan dengan lipase komersial AK Amano. Tujuan penelitian ini adalah mendapatkan mutan kapang dengan aktivitas transesterifikasi yang lebih tinggi dibandingkan tipe liarnya (KB). Proses mutasi dilakukan dengan menggunakan sinar ultraviolet (UV), ethyl methane sulfonate (EMS), dan N-methyl-N’-nitro-N-nitrosoguanidine (NMNG) terhadap kapang KB. Mutasi KB dengan menggunakan sinar UV menghasilkan 11 isolat, dimana isolat dengan kode m4.1KB1 menghasilkan aktivitas transesterifikasi yang lebih tinggi dibandingkan tipe liar, yaitu 0,172 U·mg-1. Mutan m5.7KB, yang dihasilkan dari mutan m4.1KB1 dengan perlakuan EMS, mengalami penurunan aktivitas transesterifikasi hingga hanya sebesar 0,051 U·mg-1. Mutan m6.0,3KB2 hasil perlakuan NMNG mengalami peningkatan aktivitas transesterifikasi sebesar 91,2% lebih tinggi dari KB.Kata Kunci: ethyl methane sulfonate, kapang mutan, lipase, N-methyl-N’-nitro-N-nitrosoguanidine, ultraviolet
PENGARUH TEKNIK PENGAPUNGAN HAYATI MELALUI FERMENTASI Rhizopus sp. TERHADAP KANDUNGAN NUTRISI PAKAN IKAN APUNG Paramadini, Sabrina Ayu; Sriherwanto, Catur; Nurlaila, .; Suja'i, Imam; Annisa, Mutiara Ayu
Jurnal Bioteknologi & Biosains Indonesia (JBBI) Vol 6, No 1 (2019): June 2019
Publisher : Badan Pengkajian dan Penerapan Teknologi (BPPT)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (756.154 KB) | DOI: 10.29122/jbbi.v6i1.3477

Abstract

Influence of Biofloatation Technique through Rhizopus sp. Fermentation on the Nutritional Quality of the Floating Fish FeedABSTRACTSolid fermentation using the mold Rhizopus has been used as an alternative method to improve the physical quality of fish feed, namely stability in water and floatability. Although the fermented fish feed produced had been shown previously to have better stability in water and floatability, the effect of the fermentation on the fish feed nutrition has not yet been known. This study aimed to determine the effect of solid fermentation using the mold Rhizopus sp. on the nutrient content of the fermented feed. In addition, the effect of adding tapioca as much as 0, 1, 2, 3, and 4 g to the dry weight loss and density of the fermented feed was investigated. The results showed that the fermented feed contained higher levels of ash and protein than that before fermentation. The addition of tapioca up to 4 g had no significat effect (p>0.01) on the dry weight loss of the fermented feed, but tended to increase its density compared to those without tapioca addition.Keywords: density, fermentation, fish feed, nutrition, Rhizopus ABSTRAKFermentasi padat menggunakan kapang Rhizopus telah digunakan sebagai metode alternatif untuk memperbaiki kualitas fisik pakan ikan, yakni stabilitas dalam air dan daya apung. Meskipun pakan ikan fermentasi yang dihasilkan sebelumnya sudah dibuktikan memiliki stabilitas dalam air dan daya apung yang lebih baik, namun pengaruh fermentasi terhadap nutrisi pakan ikan tersebut belumlah diketahui. Penelitian ini bertujuan mengetahui pengaruh fermentasi padat menggunakan kapang Rhizopus sp. terhadap kandungan nutrisi pakan fermentasi, dan pengaruh penambahan tapioka sebanyak 0, 1, 2, 3, dan 4 g terhadap kehilangan berat kering dan mass jenis pakan hasil fermentasi. Hasilnya menunjukkan bahwa pakan fermentasi mengandung kadar abu dan protein lebih tinggi dibandingkan sebelum difermentasi. Penambahan tapioka hingga 4 g tidak berpengaruh nyata (p>0,01) pada kehilangan berat kering pakan fermentasi, namun cenderung meningkatkan massa jenis dibandingkan tanpa penambahan tapioka.Kata kunci: fermentasi, massa jenis, nutrisi, pakan ikan, Rhizopus
PENGARUH VARIASI KONSENTRASI METANOL DAN LAMA INDUKSI TERHADAP EKSPRESI PROINSULIN OLEH Pichia pastoris SECARA INTRASELULER Martius, Efrida; Triyadi, Andree; Sofia, Dewi Yustika; Mahsunah, Anis Herliyati
Jurnal Bioteknologi & Biosains Indonesia (JBBI) Vol 6, No 1 (2019): June 2019
Publisher : Badan Pengkajian dan Penerapan Teknologi (BPPT)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1800.186 KB) | DOI: 10.29122/jbbi.v6i1.3176

Abstract

The Effects of Variation in Methanol Concentration and Induction Time on Intracellular Proinsulin Expression by Pichia pastoris ABSTRACTDiabetes is a metabolic disorder characterized by hyperglycemia. There were 215 million diabetic patients in 2014 and the number is expected to rise in 2040. Generally, insulin is used to treat diabetic patients. Insulin production by recombinant technology has been done, though still inefficient, by using E. coli and S. cerevisiae expression system. Another alternative expression system is methylotrophic yeast Pichia pastoris. In this research, proinsulin has been expressed by P. pastoris intracellularly. P. pastoris strains used in this research were X33, GS115, and KM71H. All recombinant strains were MutS. Best cultivation media was BMGY. Proinsulin expression was observed at 25°C. Pichia pastoris strain that expressed proinsulin best was GS115-PI. It was supported by PCR in which the strain GS115-PI gave 504 bp-sized bands. Based on proinsulin formation time, the final methanol concentration of 0.5% in 72 hours was found to be the best treatment.Keywords: BMGY, methanol, phenotype, Pichia pastoris, proinsulin ABSTRAKDiabetes melitus merupakan kelainan yang ditandai dengan hiperglikemia. Penderita diabetes pada tahun 2014 di dunia mencapai 215 juta dan diperkirakan akan meningkat pada tahun 2040. Umumnya penderita diabetes diberi pengobatan insulin sehingga menunjukkan akan ada peningkatan kebutuhan insulin. Produksi insulin dengan teknologi DNA rekombinan telah dilakukan dengan menggunakan sistem ekspresi E. coli dan S. cerevisiae namun masih belum efisien. Sistem alternatif lain adalah ragi metilotropik Pichia pastoris. Dalam penelitian ini dilakukan ekspresi proinsulin dari P. pastoris secara intraseluler. Galur P. pastoris yang digunakan dalam penelitian ini adalah X33, GS115, dan KM71H. Semua galur rekombinan adalah MutS. Media tumbuh terbaik adalah BMGY. Ekspresi proinsulin terlihat pada suhu 25°C. Hasil PCR menunjukkan bahwa galur GS115-PI yang dapat menghasilkan pita amplikon berukuran 504 bp. Hasil PCR ini dibuktikan oleh hasil seleksi galur yang menunjukkan bahwa galur GS115-PI dapat mengekspresi proinsulin dibandingkan galur lainnya. Berdasarkan kecepatan pembentukan pita protein proinsulin, variasi konsentrasi akhir metanol 0,5% dengan lama induksi 72 jam merupakan perlakuan terbaik.Kata Kunci: BMGY, fenotipe, metanol, Pichia pastoris, proinsulin
EFFECT OF TROGLITAZONE ON MORPHOLOGICAL CHANGES AND PPAR-γ GENE EXPRESSION IN 3T3-L1 ADIPOCYTE CELLS Sulfianti, Asri; Triwulansari, Mayriska; Nuralih, .; Churiyah, .
Jurnal Bioteknologi & Biosains Indonesia (JBBI) Vol 6, No 1 (2019): June 2019
Publisher : Badan Pengkajian dan Penerapan Teknologi (BPPT)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (820.23 KB) | DOI: 10.29122/jbbi.v6i1.3117

Abstract

Efek Troglitazone terhadap Perubahan Morfologi dan Ekspresi Gen PPAR- γ di Dalam Sel Adiposa 3T3-L1 ABSTRACT3T3-L1 cells are extensively used as a model to study adipogenesis. However, one major concern is the prolonged period of time it takes the cells to differentiate into adipocytes form. To induce this differentiation, the adipogenic induction media is required. In this study, troglitazone, a hypoglycemic agent was added to adipogenic induction media and observed in order to determine the morphological changes and peroxisome proliferator-activated receptor gamma (PPAR-γ) gene expression in 3T3-L1 differentiation. It is generally known that PPAR-ꝩ plays an important role as a transcription factor in adipocyte differentiation. Based on Oil Red O Staining, adipogenic induction with or without troglitazone changed the 3T3-L1 pre-adipocytes into mature round fat cells characterized by red droplet lipids. This cell also had a high absorbance level and degree of droplet accumulation of P≤ 0.05 in each group. In addition, cells treated by troglitazone had the highest PPAR-ꝩ mRNA level (1.9 fold) than those treated by adipogenic induction media without troglitazone or cells un-treated at all. Keywords: 3T3-L1, adipocyte, differentiation, PPAR-ꝩ, troglitazone ABSTRAKSel 3T3-L1 adalah jenis sel yang banyak digunakan dalam studi adipogenesis. Namun, salah satu kelemahan sel tersebut adalah lamanya waktu yang dibutuhkan bagi sel pre-adiposa untuk berdiferensiasi menjadi sel adiposa. Selain itu, dibutuhkan pula media induksi khusus untuk mengubah sel menjadi sel adiposa. Pada penelitian ini, kami mengobservasi fungsi troglitazone, sebagai antidiabetes terhadap perubahan morfologi dan ekspresi gen peroxisome proliferator-activated receptor gamma (PPAR-γ). Telah diketahui bahwa PPAR-ꝩ berperan penting sebagai factor transkripsi dalam diferensasi sel adiposa. Berdasarkan pewarnaan ORO, induksi sel pre-adiposa 3T3-L1 dengan media induksi dengan dan tanpa troglitazone merubah sel preadiposa menjadi sel berbentuk bulat yang dikarakterisasi dengan akumulasi droplet lemak. Nilai absorbansi sel adiposa juga menandakan adanya perbedaan yang signifikan antara kelompok sel yang diberi troglitazone dan tidak, dan sel tanpa diberi media induksi. Sementara, pada kelompok sel yang diberi troglitazone memiliki ekspresi mRNA PPAR-ꝩ (1,9 kali) tertinggi jika dibandingkan dengan sel yang diberi media induksi tanpa troglitazone, dan tanpa media induksi sama sekali.Kata Kunci: 3T3-L1, adiposa, diferensiasi, PPAR-ꝩ, troglitazone
PERBANDINGAN TIGA KIT EKSTRAKSI RNA UNTUK ANALISIS TRANSKRIPTOMIKA PADA KELAPA SAWIT (Elaeis guineensis Jacq.) Zulaeha, Siti; Purwoko, Devit; Cartealy, Imam; Tajuddin, Teuku; Karyanti, .; Khairiyah, Hayat
Jurnal Bioteknologi & Biosains Indonesia (JBBI) Vol 6, No 1 (2019): June 2019
Publisher : Badan Pengkajian dan Penerapan Teknologi (BPPT)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (143.283 KB) | DOI: 10.29122/jbbi.v6i1.3372

Abstract

Comparison of Three RNA Extraction Kits for Transcriptome Analysis of Oil Palm (Elaeis guineensis Jacq.) ABSTRACTObtaining high-quality RNA is very important at an early stage of molecular biology research. To isolate RNA, high skill and caution are required in following laboratory procedures because RNA is easily degraded, especially samples from plant tissue culture. One of the parameters used to check the total RNA quality is RIN (RNA Integrity Number). The aim of this study was to obtain RNA extraction methods on oil palm leaves, callus and somatic embryos that were of good quality and high concentrations for transcriptomic analysis. RNA extraction was carried out using Plant RNA PureLink (Ambion), Genezol RNA Extraction (Geneaid) and RibospinTM Plant (Geneall) kit methods. The results showed that oil palm leaf, callus and somatic embryo RNA were successfully extracted using the RibospinTM (Geneall) kit. Based on the total RNA number of more than 4 μg and the RIN value of more than 7, the extracted RNA could be used in RNA sequencing for transcriptomic analysis. Keywords: callus, oil palm, RNA analysis, RNA quality, somatic embryo ABSTRAKMenghasilkan RNA berkualitas tinggi sangatlah penting pada tahap awal penelitian biologi molekuler. Untuk mengisolasi RNA diperlukan keterampilan dan kehati-hatian tinggi dalam mengikuti prosedur di laboratorium karena RNA lebih mudah terdegradasi, khususnya sampel hasil kultur jaringan tanaman. Salah satu parameter yang digunakan pada pengecekan kualitas RNA total adalah RIN (RNA Integrity Number). Penelitian bertujuan mendapatkan metode ekstraksi RNA pada daun, kalus dan embrio somatik kelapa sawit yang berkualitas baik dan memiliki konsentrasi tinggi untuk analisa transkriptomika.  Ekstraksi RNA dilakukan menggunakan metode kit Plant RNA PureLink (Ambion), Genezol RNA Extraction (Geneaid) dan RibospinTM Plant (Geneall). Hasil menunjukkan bahwa RNA daun, kalus dan embrio somatik kelapa sawit telah berhasil diekstraksi dengan menggunakan kit RibospinTM (Geneall). RNA hasil ekstraksi tersebut dapat digunakan untuk sekuensing RNA dengan tujuan analisis transkriptomika, dilihat dari jumlah total RNA yang lebih dari 4 μg dan nilai RIN lebih dari 7. Kata Kunci: analisis RNA, embrio somatic, kalus, kelapa sawit, kualitas RNA 
METODE EKSTRAKSI DNA TANAMAN TANPA PRESIPITASI ETANOL UNTUK KEGIATAN POLYMERASE CHAIN REACTION (PCR) Nugroho, Kristianto; Terryana, Rerenstradika Tizar; Reflinur, .; Lestari, Puji
Jurnal Bioteknologi & Biosains Indonesia (JBBI) Vol 6, No 1 (2019): June 2019
Publisher : Badan Pengkajian dan Penerapan Teknologi (BPPT)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (196.537 KB) | DOI: 10.29122/jbbi.v6i1.3082

Abstract

A Simplified Plant DNA Extraction Protocol without Ethanol Precipitation for Polymerase Chain Reaction (PCR) Activities ABSTRACTMolecular-based research in agriculture includes DNA extraction stage involving DNA precipitation using ethanol or isopropanol which tends to take a long time. The purpose of this study was to obtain a plant DNA extraction method for Polymerase Chain Reaction (PCR) activities without going through the ethanol precipitation stage. Five important agricultural commodity crops, namely rice, corn, soybeans, chilies, and shallots were extracted by DNA using the modified Doyle and Doyle method. After the extraction phase using chloroform and isoamil alcohol solvents, the supernatant obtained was not precipitated using ethanol but was directly diluted and used as a template in PCR activities using two pairs of Simple Sequence Repeat (SSR) markers. The results showed that all samples could be well amplified, and amplicon tape visualized in both 1% agarose gel and 6% polyacrylamide gel were clearly visible. This method could save time and material, and reduce the dependence on liquid nitrogen. But this method is still limited to PCR requirements only, and cannot be used for activities that require high quality and quantity of DNA such as Next Generation Sequencing (NGS), digestion, and hybridization.Keywords: DNA extraction, ethanol precipitation, liquid nitrogen, PCR, SSR,  ABSTRAKPenelitian berbasis molekuler pada bidang pertanian mencakup tahapan ekstraksi DNA yang melibatkan presipitasi DNA menggunakan etanol atau isopropanol yang cenderung memakan waktu lama. Tujuan penelitian ini adalah untuk memperoleh metode ekstraksi DNA tanaman untuk kegiatan Polymerase Chain Reaction (PCR) tanpa melalui tahapan presipitasi etanol. Lima tanaman komoditas pertanian penting yaitu padi, jagung, kedelai, cabai, dan bawang merah diekstraksi DNA-nya menggunakan metode Doyle and Doyle yang dimodifikasi. Setelah tahap ekstraksi menggunakan pelarut kloroform dan isoamil alkohol, supernatan yang terbentuk tidak dipresipistasi menggunakan etanol melainkan langsung diencerkan dan digunakan sebagai template dalam kegiatan PCR menggunakan dua pasang marka Simple Sequence Repeat (SSR). Hasil menunjukkan bahwa seluruh sampel dapat teramplifikasi dengan baik serta pita hasil amplikon yang tervisualisasi baik pada gel agarosa 1% maupun gel poliakrilamid 6% terlihat jelas. Metode ini dapat menghemat waktu dan bahan serta mengurangi ketergantungan pemakaian nitrogen cair. Tetapi metode ini masih terbatas hanya untuk kebutuhan PCR saja dan tidak dapat digunakan untuk kegiatan yang membutuhkan DNA dengan kualitas serta kuantitas tinggi seperti Next Generation Sequencing (NGS), digesti, maupun hibridisasi.Kata Kunci: ekstraksi DNA, nitrogen cair, PCR, presipitasi etanol, SSR
EKSTRAK KAYU TEGERAN (Cudrania javanensis Trécul) SEBAGAI ANTI JAMUR Peniophora sp. Ilyas, Muhammad; Rosyida, Vita Taufika; Pratiwi, Diah; Indrianingsih, A. Wheni; Hernawan, .; Apriyana, Wuri; Darsih, Cici
Jurnal Bioteknologi & Biosains Indonesia (JBBI) Vol 6, No 1 (2019): June 2019
Publisher : Badan Pengkajian dan Penerapan Teknologi (BPPT)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (665.684 KB) | DOI: 10.29122/jbbi.v6i1.3168

Abstract

Antifungal Activity of Tegeran Wood (Cudrania javanensis Trécul) Extracts against Fungus Peniophora sp.  ABSTRACTThe fungus Peniophora sp. can degrade natural dyes, causing discoloration on batik cloth. This study was aimed to evaluate the antifungal activity of tegeran wood (Cudrania javanensis) extracts against Peniophora sp. isolated from the aqueous extract of mahogany (Swietenia mahagoni) bark. Aqueous and methanol extraction procedures gave tegeran wood extracts whose concentrations were then varied into 100, 250 and 500 ppm. Phytochemical analysis of the extracts were carried out, and the results showed that the average values of the total polysaccharides and terpenoids of both aqueous and methanol extracts did not differ significantly (p<0.05), whereas those of polyphenols did (p<0.05). The total polyphenols of the methanol extract (484.723 mg GAE/g) was higher than that of the aqueous extract (389.903 mg GAE/g). Results showed that the methanol extract of tegeran wood showed higher antifungal activity against Peniophora sp. than the aqueous extract, and was also higher than ketoconazole control (100 ppm). The minimum inhibitory concentration of the methanol extract against Peniophora sp. growth was of 500 ppm with the antifungal activity value (AFA) of 76.30%.Keywords: antifungal activity, Cudrania javanensis, Peniophora sp., Swietenia mahagoni, wood extract ABSTRAKJamur Peniophora sp. dapat mendegradasi zat warna alam sehingga warna yang dihasilkan pada kain batik lebih pudar. Penelitian ini dilakukan untuk mempelajari aktivitas anti jamur dari ekstrak kayu tegeran (Cudrania javanensis) terhadap Peniophora sp. yang diisolasi dari ekstrak air kulit mahoni (Swietenia mahagoni). Ekstraksi menggunakan metanol dan air, menghasilkan ekstrak kayu tegeran, yang kemudian konsentrasinya divariasi menjadi 100, 250 dan 500 ppm. Analisis fitokimia ekstrak kayu tegeran dilakukan, dan hasilnya menunjukkan bahwa nilai rata-rata total polisakarida dan terpenoid ekstrak metanol dan air kayu tegeran tidak berbeda nyata (p<0,05). Sedangkan total polifenol kedua ekstrak berbeda nyata (p<0,05). Total polifenol ekstrak metanol kayu tegeran (484,723 mg GAE/g) lebih besar dibandingkan dengan ekstrak air (389,903 mg GAE/g). Hasil menunjukkan bahwa aktivitas anti jamur ekstrak metanol lebih baik dibandingkan dengan ekstrak air kayu tegeran, dan juga lebih tinggi dibandingkan dengan kontrol ketoconazole (100 ppm). Konsentrasi minimum ekstrak metanol kayu tegeran yang dapat menghambat pertumbuhan jamur Peniophora sp. adalah 500 ppm dengan nilai penghambatan (AFA) sebesar 76,30%.Kata Kunci: aktivitas anti jamur, Cudrania javanensis, ekstrak kayu, Peniophora sp., Swietenia mahagoni

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