Perbandingan Kuantitas dan Kualitas DNA Bacillus sp. antara Heat Treatment dan Filter berbasis Kit The Quantity and Quality Comparison of Bacillus sp. DNA between Heat Treatment and Filter Based Kit

Main Article Content

Durrotun Ni'mah
Muhammad Zainul Fadli
Rio Risandiansyah


Diagnosis of bacterial infection can be done quickly with simplified DNA isolation methods, such as Heat Treatment, which is a simple and inexpensive DNA isolation method, but detection and quantification levels of Bacillus sp. still unknown. This study aims to compare the quantity and quality of DNA isolates from Bacillus sp. using Heat Treatment etraction method and Filter Based isolation method. This research used in vitro experiment method. Heat Treatment is compared to Filter Based Kit according to Limit of Detection (LoD) and Limit of Quantification (LoQ). Data analyzed with ANOVA and Honesty Significant Difference (HSD). DNA yield obtained by Heat Treatment at the highest concentration was 5 µg/ml, whereas Filter Based Kit was 3.67 µg/ml. LoD of the Heat treatment and Filter Based Kit were 1.61 and 5.72 µg/ml, while LoQ of the Heat treatment and Filter Based Kit were 4.87 and 17.34 µg/ml. The number of bacteria that exceeded the LoD and LoQ of Heat Treatment was 101 and 104 CFU/ml, whereas the LoD and LoQ of Filter Based Kit was >104 CFU/ml. The quantitiy of Bacillus sp. DNA with Heat Treatment method is better than Filter Based Kit. The quality of Bacillus sp. DNA both the Heat Treatment and Filter Based Kit methods are not good. 

Keywords: DNA isolation, Bacillus sp., heat treatment, Filter Based Kit


Penegakkan diagnosis infeksi bakteri dapat dilakukan secara cepat dengan metode isolasi DNA yang telah disederhanakan, seperti Heat Treatment yang merupakan metode ekstraksi DNA sederhana dan ekonomis, namun tingkat deteksi dan kuantifikasi terhadap Bacillus sp. masih belum diketahui. Penelitian ini bertujuan membandingkan kuantitas dan kualitas DNA dari Bacillus sp. dengan menggunakan metode ekstraksi Heat Treatment dan metode isolasi Filter Based Kit. Penelitian ini menggunakan metode penelitian eksperimen secara in vitro. Heat Treatment dibandingkan dengan Filter Based Kit berdasarkan Limit of Detection (LoD) and Limit of Quantification (LoQ). Analisa data menggunakan uji ANOVA serta uji Honesty Significant Diference (HSD). Yield DNA yang didapatkan Heat Treatment pada konsentrasi terbesar adalah 5 µg/ml, sedangkan dengan Filter Based Kit adalah 3.67 µg/ml. Nilai LoD Heat treatment dan Filter Based Kit adalah 1.61 µg/ml dan 5.72 µg/ml, sedangkan nilai LoQ Heat treatment dan Filter Based Kit adalah 4.87 µg/ml dan 17.34 µg/ml. Jumlah bakteri yang melebihi nilai LoD dan LoQ Heat treatment adalah 101 CFU/ml dan 104 CFU/ml, sementara LoD dan LoQ Filter Based Kit adalah >104 CFU/ml. Kuantitas DNA Bacillus sp. dengan metode Heat Treatment lebih baik dibandingkan dengan Filter Based Kit. Kualitas DNA Bacillus sp. baik dengan metode Heat Treatment maupun Filter Based Kit tidak baik.

Kata kunci: isolasi DNA, Bacillus sp., heat treatment, Filter Based Kit

Article Details

How to Cite
Ni’mah, D., Fadli, M., & Risandiansyah, R. (2021). Perbandingan Kuantitas dan Kualitas DNA Bacillus sp. antara Heat Treatment dan Filter berbasis Kit. BIOSAINTROPIS (BIOSCIENCE-TROPIC), 6(2), 86-95.
Article (Makalah)


[1] Lu Z, Guo W, Liu C. 2018. Isolation, identification, and characterization of novel Bacillus subtilis. Journal of Veterinary Medical Science.16-0572.
[2] Ping DY, Chao XZ, Lian YG, Wei L, Feng ZQ, Hong YD, Jie L, Li C, Yun Z, Yi XC. 2019. A newly isolated Bacillus subtilis strain named WS-1 inhibited diarrhea and death caused by pathogenic Escherichia coli in newborn piglets. Frontiers in Microbiology. 10:1248.
[3] Muthén LK, Muthen B. 2017. Mplus user's guide: Statistical analysis with latent variables, user's guide. Muthén & Muthén;
[4] Gupta N. 2019. DNA extraction and polymerase chain reaction. Journal of cytology. 36(2):116-7.
[5] Boesenberg-Smith KA, Pessarakli MM, Wolk DM. 2012 Assessment of DNA yield and purity: an overlooked detail of PCR troubleshooting. Clinical Microbiology Newsletter. 34(1):1-6.
[6] Mulyani Y, Purwanto A, Nurruhwati I. 2011. Perbandingan beberapa metode isolasi DNA untuk deteksi dini koi herpes virus (KHV) pada ikan mas (Cyprinus carpio L.). Jurnal Akuatika. 2(1).
[7] Berensmeier S. 2006. Magnetic particles for the separation and purification of nucleic acids. Applied microbiology and biotechnology. 73(3):495-504.
[8] Syafaruddin S, Randriani E, Santoso TJ. 2011. Efektivitas dan efisiensi teknik isolasi dan purifikasi DNA pada jambu mete. Journal of Industrial and Beverage Crops. 2(2):141601.
[9] Currie LA. 1995. Nomenclature in evaluation of analytical methods including detection and quantification capabilities (IUPAC Recommendations 1995). Pure and applied chemistry. 67(10):1699-723.
[10] Nutz S, Döll K, Karlovsky P. 2011. Determination of the LOQ in real-time PCR by receiver operating characteristic curve analysis: application to qPCR assays for Fusarium verticillioides and F. proliferatum. Analytical and bioanalytical chemistry. 401(2):717-26.
[11] Munch DJ, Wasko M, Flynt E, Wendelken SC, Scifres J, Mario JR, Hunt M, Gregg D, Schaeffer T, Clarage M, Lumpkin MS. Validation and Peer Review of US Environmental Protection Agency Chemical Methods of Analysis.
[12] Sains P. nd. Validasi Metode Untuk Analisis Kandungan Uranium Menggunakan Potensiometer T-90. Metode.;3:4.
[13] Food and Drug Administration. Bacteriological analytical manual. revision A.
[14] Mayanti B, Ariesyady H. 2009. Identifikasi Keberagaman Bakteri pada Commercial-Seed Pengolah Limbah Cair Cat. Jurnal Teknik Lingkungan. 16(1):52-61.
[15] Murtiyaningsih H. 2017. Isolasi DNA genom dan identifikasi kekerabatan genetik nanas menggunakan RAPD (Random Amplified Polimorfic DNA). Agritrop: Jurnal Ilmu-Ilmu Pertanian (Journal of Agricultural Science). 15(1).
[16] Mulyatni AS, Budiani A, Taniwiryono D. nd. Aktivitas antibakteri ekstrak kulit buah kakao (Theobroma cacao L.) terhadap Escherichia coli, Bacillus subtilis, dan Staphylococcus aureus. E-Journal Menara Perkebunan. 80(2).
[17] BIOL 3716: Molecular Biology I. Protocol: Spectrophotometric Determination of DNA Quantity and Purity. Genetics & Biotechnology Lab. v091708. Diterima Tanggal 23 Agustus 2020. URL:
[18] Ahmed OB, Asghar AH, Elhassan MM. 2014. Comparison of three DNA extraction methods for polymerase chain reaction (PCR) analysis of bacterial genomic DNA. African Journal of Microbiology Research. 8(6):598-602.
[19] Hidayani AA, Tassakka AC, Parenrengi A. 2016. Isolation and characterization of an envelope protein (VP19) of a White Spot Syndrome Virus from diseased vannamei (Litopenaeus vannamei) in Indonesia. Aquaculture, Aquarium, Conservation & Legislation. 9(2):389-95.
[20] Riahi M, Babaei M, Ghahremaninejad F. 2019. Genomic DNA Isolation From Scrophularieae Dried Leaves Using A Simple, High-Throughput Protocol. Bangladesh Journal Of Botany. 48(4):1231-5.
[21] Safitri R, Muchlissin SI, Mukaromah AH, Darmawati S, Ethica SN. 2018. Isolasi Bakteri Penghasil Enzim Protease Bacillus Thuringiensis Irodi Pada Oncom Merah Pasca Fermentasi 24 Jam. InProsiding Seminar Nasional & Internasional (Vol. 1, No. 1).
[22] Shi R, Lewis RS, Panthee DR. 2018. Filter paper-based spin column method for cost-efficient DNA or RNA purification. Plos one. 13(12):e0203011.
[23] Cheng HR, Jiang N. 2006. Extremely rapid extraction of DNA from bacteria and yeasts. Biotechnology letters 28(1):55-9.
[24] Esser KH, Marx WH, Lisowsky T. 2006. maxXbond: first regeneration system for DNA binding silica matrices. Nature methods. 3(1):i-i.
[25] Melzak KA, Sherwood CS, Turner RF, Haynes CA. 1996. Driving forces for DNA adsorption to silica in perchlorate solutions. Journal of colloid and interface science. 181(2):635-44.
[26] Radji M. 2006. Penuntun Praktikum Mikrobiologi Farmasi. Edisi ke 2. Departemen Farmasi FMIPA-UI. hal15-43.