Detection of porcine contamination in food material by employing PCR techniques is integral in halal food confirmation. However, PCR is both costly and laborious, particularly in DNA isolation method. This study explores several different methods in DNA extraction for PCR amplification in bovine and porcine raw and boiled meat samples. Four methods for DNA extraction (conventional PCI method, DNA isolation kit, alkaline-based method, and a DNA lysis buffer-only from the same kit) was employed followed by PCR using primers from previous studies and compared for DNA quality and quantity (in six replicates) and PCR amplification on the best three DNA samples. This study shows that in all samples, the conventional method had the best DNA yield based on nanodrop measurement, followed by an alkali-based method, buffer-only method, and DNA isolation kit. Each method except lysis-buffer only had at least one sample with good DNA quality. Conventional and isolation kit showed reliable positive PCR detection for all porcine and bovine samples (92% positive). Using the alkaline-lysis method, DNA was amplified reliably on boiled meat samples (83% positive). Lysis-buffer-only method did not show consistent PCR amplification on the samples used (50% positive). The conclusion was that conventional PCI method and DNA isolation kit showed high reliability in PCR amplification of bovine and porcine meats, both raw and boiled. While high DNA yield was obtained using the alkaline-lysis method, PCR amplification was only successful on boiled samples. Lysis-buffer only method yielded in poor DNA quality and was not able to result in reliable DNA amplification.

Alkaloid merupakan senyawa yang terkandung dalam Imperata cylindrica L. (alang – alang). Ekstrak metanolik I. cylindrica L. diketahui berinteraksi dengan beberapa jenis antibiotik. Namun, belum ada penelitian yang membuktikan bahwa alkaloid dari I.cylindrica L. dapat meningkatkan kinerja antibiotik.Penelitian ini mengisolasi senyawa alkaloid dan melihat pengaruh penambannya terhadap zona hambat amoksisilin dan kloramfenikol terhadap Staphylococcus aureus.Metode: Ekstraksi dilakukan dengan maserasi menggunakan kloroform dan soxhletasi menggunakan methanol. Fraksinasi dengan pelarut, yaitu aquadest, methanol dan etil asetat. Uji fitokimia secara kualitatif dengan melihat perubahan warna. Uji Zone of Inhibition (ZOI) dilakukan untuk mengetahui efek dari kombinasi fraksi alkaloid I.cylindrica L. dengan antibiotik terhadap S.aureus dengan metode Kirby-Bauer. Diameter zona bening diukur menggunakan jangka sorong, dan interpretasi hasil berdasarkan metode Ameri-Ziaei Double Antibiotic Synergism Test (AZDAST).Hasil : Fraksi – fraksi alkaloid I.cylindrica L. tunggal tidak membentuk zona bening terhadap S.aureus. Kombinasi fraksi alkaloid dengan kloramfenikol (F1C dan F2C) memiliki ZOI dengan rerata diameter 30.73 ± 0.9 mm dan 30.53 ± 0.55 mm, yang lebih besar dari ZOI kloramfenikol tunggal yaitu 28.6 ± 0.95 mm dan fraksi tunggal 0 ± 0 mm. Pada uji fitokimia ditemukan bahwa senyawa yang terkandung adalah alkaloid.Simpulan: Alkaloid merupakan senyawa aktif dari I.cylindrica L.. Fraksi alkaloid I.cylindrica L. bersifat potensiasi dengan antibiotik kloramfenikol terhadap S.aureus karena mampu meningkatkan kinerja antibiotik tersebut.

No abstract

One of housekeeping genes is actin gene. This gene is frequently used in gene expression studies as an internal control. The DNA sequence encoding actin from Pandan (Benstonea sp.) clone Riau has not been reported, therefore, this study investigated the DNA sequence encoding actin isolated from Benstonea sp. clone Riau. Total DNA isolation was performed in fresh leaves, total RNA isolation from stem, total cDNA synthesis, polymerase chain reaction using degenerate actin primer, electrophoresis, cloning, transformation, blue white colony selection, colony PCR, sequencing, data analysis using BioEdit and MEGA6 softwares and BLASTn program. The partial DNA sequence encoding actin from Benstonea sp. clone Riau obtained was 1,403 bp. The sequence was grouped as part of actin1 (ACT1) and it was consisted of two exons and one intron. The predicted coding and peptide sequences were 616 bp and 205 amino acids, respectively. The predicted coding sequence had 90% similarity to some ACT1 mRNA from some plants species but none of which belongs to Benstonea genus or Pandanaceae family. The deduced peptide sequence had similarity to some ACT1 peptide from some plant species of up to 99% and also none of them belongs to Benstonea genus or Pandanaceae family. Thus, the partial ACT1 gene obtained in this study was the first sequence reported from Benstonea genus. Furthermore, this sequence can be used as a reference to isolate actin genes from other species within Benstonea genus for gene expression analysis purposes.