Rancangan Primer Spesifik Gen Macrophage Mannose Receptor (MMR) Untuk Polymerase Chain Reaction (PCR) dan Sekuensing Deoxyribo Nucleic Acid (DNA)  

Oleh:

Triyani, Yani ; Nafsi, Nurizzatun ; Yuniarti, Lelly ; Sekarwana, Nanan ; Sutedja, Endang ; Gurnida, Dida Ahmad ; Parwati, Ida ; Alisjahbana, Bachti

Jenis: Article from Journal - ilmiah nasional - terakreditasi DIKTI
Dalam koleksi: Indonesian Journal of Clinical Pathology and Medical Laboratory vol. 22 no. 02 (Mar. 2016), page 158-162.
Topik: Specific Primer Design; Macrophage Mannose Receptor Gene; Polymerase Chain Reaction; Deoxyribo Nucleic Acid Sequencing; Rancangan Primer Spesifik; Gen Macrophage Mannose Receptor; Sekuensing Deoxyribo Nucleic Acid
Fulltext: I01 v22 n2 p158 kelik2017.pdf (312.49KB)

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Perpustakaan FK Nomor Panggil: I01.K Non-tandon: 1 (dapat dipinjam: 0) Tandon: tidak ada

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Isi artikel

The order (sequencing) determinationof DeoxyribonucleicAcid (DNA) bases is the gene’s most basic information, using the method of Polymerase Chain Reaction (PCR) as its stage. A key factor of successful detection by PCR is specific PCR primer design choice. The detection of diversity of Mycobacterium Mannose Receptor (MMR) gene, responsible for recognizing mannosylated antigen structure of Mycobacterium tuberculosis (M.tb) by DNA sequencing of exon 7 chromosome 10p12, related to susceptiblity for Pulmonary Tuberculosis(TB), was first performed in China in 2012. The purpose of this study was to find specific primerfromboth design originated from the research in China/primer I and my own design/primer IIby using Primer3 software. This study was based on 10 healthy subjects and was a preliminary study of a research titled. The Relationship of Single Nucleotide Polymorphisms (SNPs) of Macrophage Mannose Receptor Gene to Pulmonary Tuberculosis Cases. The examination materials consist of 3 mL of EDTA blood and DNA extraction from its buffy coat. The resulting DNA was processed by PCR to amplify MMR gene with primer I and II. The primer I successfully amplified DNA fragments up to 780bp while primer II only 329 bp. The MMR gene DNA sequencing analysis was performed on the amplification result of both kinds primers by using DNA Baser and Ensembl-BLAST software. The results were different, DNA sequencing result by using the primer I was found in several chromosomes and also in several loci. Whereas, by using the primer II, it was only found in chromosome 10 and in the same locus. Based on this study, it can be concluded that the specific primer design is one of the most important factors in the success of DNA sequencing.

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