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Low cost and comprehensive pork detection in processed food products with a different food matrix

Low cost and comprehensive pork detection in processed food products with a different food matrix

Fenny Aulia Sugiana, Henni Widyowati, M. Ali Warisman, Suryani, and Desriani (2018)

Indonesian Journal of Biotechnology, 23 (1): 21-27.ISSN 0853-8654

 Low cost and comprehensive pork detection in processed food products with a different food matrixThe adulteration of processed beef-based meat products with pork is a sensitive issue in Indonesia. Therefore a simple, low cost, and accurate method is required for the detection of pork, so as to protect consumers from accidental consumption of adulterated meat. In this study, we developed a detection method for the low cost identification of pork in processed meat products. We used the cost-efficient Taq DNA polymerase, DreamTaq Green PCR master mix (2x), and duplex PCR method to recognize pork simultaneously with 18S rRNA detection. A positive control containing a pork gene inserted into pGEM®-T easy was prepared, along with a negative control. The results of the duplex PCR were used to assess its specificity, detection limit, and its ability to recognize pork in processed meat products with a different food matrix. 18S rRNA detection was for confirming DNA integrity of DNA extracted from the processed food, while the positive control confirmed that the reagents were working well and the negative control confirmed a non-contamination problem. Following this, the duplex PCR was opধmized and the optimum concentration primer for duplex PCR detection was found to be 3 µM for pork and 0.2 µM for 18S rRNA. As liħle as 3.125 ng of the DNA template could be used to detect whether a sample contained pork. Of the nine commercial processed meat products tested, five were found to contain pork while four halal products showed no signs of pork. It can be concluded that duplex PCR is a simple, fast, sensitive, specific, and low cost method of detecting pork in processed meat products.

Keywords: 18S rRNA, duplex PCR, low cost, pork, processed meat

Katalog: http://perpus.biotek.lipi.go.id/index.php?p=fstream&fid=3480&bid=16089

Disusun oleh: Ludya AB/Pustakawan

 

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Tandem Recombinant Plasmid Construction as Positive Control for PIK3CA H1047R Detection Based on SYBR Green I qPCR

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.

 Tandem Recombinant Plasmid Construction as Positive ControlPIK3CA 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.

Katalog: http://perpus.biotek.lipi.go.id/index.php?p=fstream&fid=3475&bid=16075

Disusun oleh: Ludya AB/Pustakawan

 

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Mahasiswa Universitas Nasional Jakarta Kunjungi Pusat Penelitian Bioteknologi LIPI

Cibinong-Humas LIPI. Pusat Penelitian Bioteknologi Lembaga Ilmu Pengetahuan Indonesia (LIPI) menjadi  salah satu objek dalam memperdalam bidang  ilmu hayati,  seperti halnya kali ini Fakultas Biologi Universitas Nasional Jakarta, mengadakan kunjungan ke Puslit Bioteknologi,  pada Senin (16/12) 2019. Rombongan yang terdiri dari dosen pendamping dan mahasiswa ini diterima di gedung Meat-Milk Pro Puslit Bioteknologi-LIPI oleh tim Humas Kawasan CSC-BG, Cibinong. Sebelum melakukan kegiatan, terlebih dahulu pengunjung mendapatkan informasi terkait kegiatan penelitian melalui tayangan video profil LIPI dan Pusat Penelitian Bioteknologi, agar pengunjung  mendapatkan informasi tentang kegiatan penelitian yang dilaksanakan oleh Puslit Bioteknologi di laboratorium.

Selanjutnya kegiatan dilanjutkan dengan kunjungan ke beberapa Laboratorium, mahasiswa dan pendamping yang berjumlah 20 orang di arahkan  untuk melihat langsung Laboratorium yang ada di Pusat Penelitian Bioteknologi-LIPI yaitu : Laboratorium Biokatalis dan Fermentasi serta Laboratorium Bioenergi dan Bioproses. Peserta kunjungan yang ke Lab Biokatalis dan Fermentasi di pandu Ade Andriani, mengutarakan bahwa tentang kegiatan pada Lab penelitian ini  dilakukan adalah  dengan memanfaatkan mikroba dan enzim melalui fermentasi/reaksi berbasis rekayasa genetika mikroba. Lanjut Ade, beberapa produk utama yang dihasilkan adalah Oligosakarida (pangan fungsional), enzim kasar dan enzim murni untuk kebutuhan industri pangan, farmasi dan energi, Penyediaan mikroba potensial untuk industri dan lingkungan serta Produk hasil fermentasi. Pada Lab Bioenergi dan Bioproses, Swastika menjelaskan tentang pengembangan bioenergi dari mikroba fotosintesa dan pengembangan total proses dari bioenergi (biorifenari dari bioenergi).

Kepada pengunjung disampaikan bahwa Pusat Penelitian Bioteknologi juga menawarkan beberapa program yang dapat diikuti oleh para mahasiswa yang berminat di bidang bioteknologi seperti mengikuti kegiatan pelatihan dan bimbingan mengingat Puslit  Bioteknologi memiliki 17 laboratorium dengan kompetensi inti yang berbeda-beda. (est/edt:ks)

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