Claim Missing Document
Check
Articles

Found 2 Documents
Search
Journal : METAMORFOSA Journal of Biological Sciences

TEKNIK PERANCANGAN PRIMER UNTUK SEKUEN GEN MDR-1 VARIAN 1199 PADA SAMPEL BUFFY COAT PASIEN ANAK DENGAN LLA Putu Desy Yustinadewi; Putu Sanna Yustiantara; Inna Narayani
Metamorfosa: Journal of Biological Sciences Vol 5 No 1 (2018)
Publisher : Prodi Magister Ilmu Biologi, Fakultas MIPA, Universitas Udayana

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.24843/metamorfosa.2018.v05.i01.p16

Abstract

Single Nucleotide Polymorphism (SNP) 1199 dapat diidentifikasi menggunakan sampel buffy coat dengan metode Polymerase Chain Reaction (PCR). Komponen- komponen yang diperlukan pada proses PCR adalah template DNA; sepasang primer, yaitu suatu oligonukleotida pendek yang mempunyai urutan nukleotida yang komplementer dengan urutan nukleotida DNA templat; dNTPs (Deoxynucleotide triphosphates); buffer PCR; magnesium klorida (MgCl2) dan enzim polimerase DNA (Handoyo dan Rudiretna, 2001). Primer sangat mempengaruhi spesifitas dan sensitivitas reaksi PCR. Rancangan suatu primer merupakan salah satu parameter penentu keberhasilan suatu proses PCR (Ebd-Elsalam, 2003). Primer untuk sekuensing gen MDR-1 variant 1199 berhasil didesain dalam kondisi terbaik. Panjang sekuen primer forward sejumlah 21 oligonukleotida dan reverse sejumlah 20 oligonukleotida dengan fragmen sebesar 225 pb.
DESAIN TAQMAN PROBE SECARA IN SILICO SEBAGAI PENDETEKSI MUTASI PADA KODON 516 GEN rpoB Mycobacterium tuberculosis UNTUK METODE REAL-TIME PCR Dek Pueteri Dewi Suryani; Putu Sanna Yustiantara; Sagung Chandra Yowani
Metamorfosa: Journal of Biological Sciences Vol 4 No 1 (2017)
Publisher : Prodi Magister Ilmu Biologi, Fakultas MIPA, Universitas Udayana

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.24843/metamorfosa.2017.v04.i01.p04

Abstract

The aim of this research was to in silico design of TaqMan probe. Design of TaqMan probe were conducted using software Clone Manager Suite 6. As a template, the rpoB gene of M. tuberculosis H37Rv (accession number U12205.1) was used. The results of this research were 8 sequences such as, R516MV-2, R516MV-3, R516MV-4, R516MV-5, R516MV-7, R516MV-8, R516MV-11, R516MV-13. These sequences were met the criteria of TaqMan probe, such as length of nucleotide (23-28 nucleotide), Tm value (72ÂșC), %GC (50-58%), runs and repeats (?4 bases), dimer structure in accordance to the requirements and does not form hairpin structures. In addition, these sequences were met labeling criteria of TaqMan probe which are including the location of G bases and the number of G-C bases in sequences. Therefore, these sequences could be labeled by FAM (reporter) at 5' end and TAMRA (quencher) at 3' end. The conclusion of this research, the sequences were met the criteria of TaqMan probe. Therefore, it could be targeted to detect mutations at codon 516 with a change of aspartic acid into valine (GAC ? GTC) by using real-time PCR method.