Tandem Recombinant Plasmid Construction as Positive Control for PIK3CA H1047R Detection Based on SYBR Green I qPCR
Nahdaturrugaisiyah, Azamris, Bugi Ratno Budiarto, I Made Artika, Desriani (2018)
Pak. J. Biotechnol., Vol. 15 (3): 735-742.
PIK3CA H1047R mutation is found in breast cancer in high frequency and its detection could be applied as prognosis and predictive factor for trastuzumab therapy. qPCR is one of the simplest and robust method for PIK3CA H1047R detection. Provision of positive control for PIK3CA H1047R detection based on qPCR will support data analysis efficiently and avoid false negative result. In this research, we constructed a tandem recombinant plasmid (pGEM-tandPIK3CA) as positive control for Tm Shift SYBR Green I qPCR-based of PIK3CA H1047R detection system by ligating wild-type and PIK3CA H1047R fragments tandemly into pGEM-T Easy. The tandem plasmid was confirmed by restriction, DNA sequencing and qPCR. As a result, pGEMtandPIK3CA has been successfully constructed and confirmed. Statistical analysis shows high repeatability and reproducibility with % CV of <25%. The main advantage of this tandem positive control is its ability to serve as positive control for both wild-type PIK3CA and PIK3CA H1047R simultaneously, therefore improving the efficiency of the detection system.
Keywords: Breast cancer, PIK3CA H1047R, qPCR, positive control.
Disusun oleh: Ludya AB/Pustakawan
Antimicrobial Activity of Selenium Nanoparticles Synthesized by Actinomycetes Isolated from Lombok Island Soil Samples
Shanti Ratnakomala, Nurul Fitri Sari, Fahrurrozi and Puspita Lisdiyanti (2018)
Jurnal Kimia Terapan Indonesia. Vol. 20 (1): 8-15. e-ISSN: 2527–7669.
A total of 98 actinomycetes were isolated from the soil and litter samples collected from the cacao and coffee plantation in Lombok Island, West Nusa Tenggara of Indonesia. These isolates were screened for their antimicrobial activity. Among 98 isolated strains, only 24 isolates showed antimicrobial activity against test microorganisms, 20.4% were active against Bacillus subtilis BTCC B-612, 14.3% against Staphylococcus aureus BTCC B-611, and 5.1% against Escherichia coli BTCC B-609. Of these 24 isolates, 3 were found to be able to grow in medium containing 3 mM Selenium oxide and the culture were changed color to red. Two of the best strains, L-155 and L-156, were selected for assessing production of Selenium nanoparticles. Bioreduction of selenium nanoparticles was confirmed by UVvisible spectrophotometer that showed the peak between 300 and 320 nm. Biosynthesized selenium nanoparticle from isolate actinomycetes L-155 and L-156 were found to have a broad spectrum of activity against the tested microorganisms Bacillus subtilis, Staphylococcus aureus, Escherichia coli, Micrococcus luteus, and Candida albicans. The study shows the rapid and eco-friendly synthesis of selenium nanoparticles from soil actinomycetes. Most of these active isolates showed to possessed antibacterial property.
Keywords: Lombok Island, actinomycetes, selenium nanoparticles, antimicrobial activity
Disusun oleh: Ludya AB/Pustakawan
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