PROGRAM STUDI TEKNOLOGI LABORATORIUM MEDIS (D-3) FAKULTAS ILMU DAN TEKNOLOGI KESEHATAN UNIVERSITAS JENDERAL ACHMAD YANI 2026

SALWA NABILAH, 2350381030
OPTIMASI SUHU ANNEALING DAN KONSENTRASI PRIMER GEN LDLR METODE POLYMERASE CHAIN REACTION (PCR)
ix + 56 halaman + 5 tabel + 6 gambar + 4 lampiran


ABSTRAK

Gen Low-Density Lipoprotein Receptor (LDLR) berperan penting dalam metabolisme kolesterol, sehingga amplifikasi gen ini memerlukan kondisi Polymerase Chain Reaction (PCR) yang optimal. Keberhasilan amplifikasi dipengaruhi oleh beberapa faktor, terutama suhu annealing dan konsentrasi primer. Penelitian ini bertujuan menentukan kondisi optimal suhu annealing dan konsentrasi primer pada amplifikasi gen LDLR. Penelitian menggunakan metode deskriptif dengan melakukan optimasi suhu annealing pada 62,0°C, 60,4°C, 57,1°C, dan 54,6°C serta konsentrasi primer 0,1 µM, 0,5 µM, dan 1,0 µM. Sampel berupa DNA hasil ekstraksi darah vena orang sehat menggunakan metode spin column. Hasil amplifikasi dianalisis melalui elektroforesis gel agarosa berdasarkan ukuran, intensitas, dan spesifisitas pita DNA. Hasil penelitian menunjukkan bahwa suhu annealing 60,4°C menghasilkan pita DNA paling jelas dan spesifik pada ukuran target 369 bp. Konsentrasi primer 0,5 µM menghasilkan pita DNA dengan intensitas paling kuat dibandingkan konsentrasi lainnya. Dengan demikian, kombinasi suhu annealing 60,4°C dan konsentrasi primer 0,5 µM merupakan kondisi PCR yang paling optimal untuk amplifikasi gen LDLR. Penelitian selanjutnya disarankan menerapkan kondisi PCR optimal tersebut untuk deteksi gen LDLR pada pasien hiperkolesterolemia.





Kata Kunci: gen LDLR, PCR, suhu annealing, konsentrasi primer

Kepustakaan: 2021–2025

STUDY PROGRAM OF MEDICAL LABORATORY TECHNOLOGY (D-3) FACULTY OF HEALTH SCIENCES AND TECHNOLOGY UNIVERSITY OF JENDERAL ACHMAD YANI 2026

SALWA NABILAH, 2350381030
OPTIMIZATION OF ANNEALING TEMPERATURE AND PRIMER CONCENTRATION FOR THE LDLR GENE USING THE POLYMERASE CHAIN REACTION (PCR)
ix + 56 pages + 5 tables + 6 figure + 4 attachment


ABSTRACT


The Low-Density Lipoprotein Receptor (LDLR) gene plays an essential role in cholesterol metabolism, and its amplification requires optimal Polymerase Chain Reaction (PCR) conditions. The success of PCR amplification is influenced by several factors, particularly annealing temperature and primer concentration. This study aimed to determine the optimal annealing temperature and primer concentration for LDLR gene amplification. A descriptive study was conducted by optimizing four annealing temperatures (62.0°C, 60.4°C, 57.1°C, and 54.6°C) and three primer concentrations (0.1 µM, 0.5 µM, and 1.0 µM). DNA samples were extracted from venous blood of healthy individuals using the spin column method. PCR products were analyzed by agarose gel electrophoresis based on the size, intensity, and specificity of the DNA bands. The results showed that an annealing temperature of 60.4°C produced the clearest and most specific DNA band at the target size of 369 bp. Primer optimization demonstrated that a concentration of 0.5 µM yielded the strongest and most specific DNA band compared to the other concentrations. Therefore, the combination of an annealing temperature of 60.4°C and a primer concentration of 0.5 µM was identified as the optimal condition for LDLR gene amplification. Future studies are recommended to apply these optimized PCR conditions for the detection of the LDLR gene in patients with hypercholesterolemia.





Keywords: LDLR gene, PCR, annealing temperature, primer concentration

Literature: 2021-2025