The use of direct PCR technique without DNA extraction in the confirmation test for Salmonella typhimurium ATCC 14028 bacteria on meatball samples was carried out in the Food and Drug molecular biology testing laboratory Administration in Gorontalo. The basis of this research is to have an impact on economic value in carrying out the confirmation test for S. typhimurium ATCC 14028, where testing is carried out conventionally, namely DNA extraction, which requires a large amount of money. Hence, it is necessary to innovate to modify the testing phase so that it is more effective and efficient. The purpose of this study was to see whether the direct PCR technique without DNA extraction can be done for the confirmation test of S. typhimurium ATCC 14028 on meatball samples. This study's sample consisted of 20 types of meatball samples spiked with S. typhimurium ATCC 14028 cultures. The method used in this study was qPCR analysis using the SYBR Green method. Data analysis was carried out based on 2 main criteria: (1) Ct analysis and (2) Tm analysis. Real-time PCR analysis results obtained Ct values in the range 14.14 - 15.20 with an average of 14.82 and Tm values 85.20 - 86.30 with an average of 85.79. Based on these data, it can be concluded that using direct PCR can be used for testing confirmation of S. typhimurium ATCC 14028 on meatball samples.
The detection of Salmonella typhimurium ATCC 14028 using real-time PCR on powdered traditional medicinal products was carried out in the microbiology and molecular biology testing laboratory of the Food and Drug Administration in Gorontalo. This research aims to provide a reference for alternative testing methods in testing the products of traditional powder preparations on the market. The sample consisted of 10 traditional powder preparations spiked with positive control of S. typhimurium ATCC 14028 phase 2. The method used in the study was real-time PCR analysis using the SYBR® Green method, while DNA isolation using the direct PCR method. Data analysis was performed by analyzing the sample's melting temperature (Tm) curve and comparing it with positive control. The results showed that S. typhimurium ATCC 14028 was detected in samples at an average Tm value of 84.18°C, with ranges of 84.0-84.5°C. For positive control, the Tm value was at 85.2°C, while for the negative control, the Tm value was not detected. Based on these data, it can be concluded that S. typhimurium ATCC 14028 in traditional medicine products powder preparations can be detected using real-time PCR.
Abstract. Sophian A, Purwaningsih R, Muindar, Igirisa EPJ, Amirullah ML. 2021. Short Communication: Analysis of purity and concentration of DNA extracted from intron patho gene-spin extraction on crab processed food product samples. Asian J Trop Biotechnol 18: 28-31. The purpose of this study was to optimize the Intron patho gene-spin DNA/RNA extraction kit on crab processed food products so that it can be used for testing samples other than viral and bacterial samples. The method used in purity and concentration analysis to analyze total DNA (yield) is the absorbance method using a nanophotometer and the method used to isolate the sample DNA is the centrifuge Column or spin column method. The research data showed that the extracted sample concentration was in the range of 15.15-15.75, with an average of 15.46. while the purity values measured at the A260/A280 wavelengths were obtained with a purity range between 2,124-2,346. The total DNA analysis (yield) results obtained an average of 755.04 with the lowest yield value of 575.50 and the highest yield value of 787.50. As a suggestion for further research, it is hoped that modification can be made to perfect the isolated DNA so that it has better purity and concentration values.
BACKGROUND: The bark of pulai (Alstonia scholaris [L] R.Br.) by the Gorontalo community is used for powder preparation (as a wedding ceremony custom), containing flavonoid compounds that function as sunscreens. Thus, the bark of pulai can be made in pharmaceutical preparations in the form of lotion dosage forms. AIM: This study aims to determine the SPF value of the lotion preparation of pulai bark extract. There are four formulas, formula F0 (lotion base), formula F1 (0.5%), formula F2 (1%), formula F3 (2%). METHODOLOGY: Physical evaluation of lotion preparations was carried out with organoleptic test parameters, spreadability, homogeneity, stability, pH. The SPF value of the lotion preparation was tested using the ultraviolet-Vis Spectrophotometry method with a wavelength of 290–400. RESULTS: The results of the physical evaluation of the organoleptic test and the homogeneity test were obtained that the lotion had a clear white color (F0), light yellow (F1), yellow (F2), dark yellow (F3), a characteristic smell of olive oil, and the four formulas had a thick and homogeneous consistency. The dispersion power of F0, F1, and F2 does not meet the requirements of the dispersion test value except F3. The three lotion formulations F1, F2, and F3 showed physical instability, except for F0 (lotion base). The pH of the lotions of the four formulas was in the range that was in accordance with the standard of topical preparations. CONCLUSION: The determination of the SPF value in the formulas F1, F2, and F3, respectively, was 3.3427 (minimum protection), 4.7752 (medium protection), and 5.0968 (medium protection).
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