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
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Abstract
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
ABSTRAK
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
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