Perbandingan Metode Isolasi DNA sebagai Templat PCR untuk Identifikasi Jenis Kelamin Cerek Jawa (Charadriius javanicus) secara Molekuler Menggunakan Primer 2550F/2718R

Yulius Wahyu Pratomo, Felicia Zahida, Pramana Yuda

Abstract


In molecular studies, extraction can be time consuming and costly (especially in large scale studies), therefore some studies examine the success rate of amplification using alternative DNA isolation methods (without extraction). This study aims to determine the most effective DNA isolation method in Javan Plover molecular sexing by comparing several methods, such as extraction kit, alkaline lysis and direct PCR.This study used filter paper (5) and EDTA stored blood (8) as samples, and both sample types would be isolated using extraction kit, alkaline lysis, and direct PCR method. Molecular sexing was carried out using 2550F/2718R primer. The results showed that DNA in filter paper and EDTA stored blood, both using kit extraction, alkaline lysis and direct PCR, methods were successfully amplified with 2550F/2718R primer which produced 621 bp sized CHD1-Z and 459 bp sized CHD1-W band. The results suggested  that extraction kit is the most effective DNA isolation method for Javan Plover molecular sexing.

 

Abstrak

Dalam studi molekuler, ekstraksi dapat memakan waktu dan biaya saat diterapkan dalam penelitian skala besar, sehingga beberapa studi mengkaji tingkat keberhasilan amplifikasi menggunakan metode isolasi DNA alternatif (tanpa ekstraksi). Penelitian ini bertujuan untuk mengetahui metode isolasi DNA paling efektif untuk molecular sexing Cerek Jawa. Penelitian ini menggunakan 5 sampel darah kertas saring, dan 8 sampel darah EDTA yang diisolasi dengan metode ekstraksi kit, alkalin lisis dan PCR langsung. Molecular sexing dilakukan dengan primer 2550F/2718R. Hasil penelitian menunjukan bahwa DNA pada sampel darah kertas saring dan EDTA, baik metode ektraksi kit, alkalin lisis dan PCR langsung, berhasil diamplifikasi dengan primer 2550F/2718R yang menghasilkan pita CHD1-Z dengan ukuran 621 bp dan pita CHD1-W dengan ukuran 459 bp. Penelitian ini menunjukan bahwa ekstraksi DNA dengan kit menghasilkan DNA template yang paling efektif untuk molecular sexing Cerek Jawa.


Keywords


Javan Plover, Molecular Sexing, 2550F/2718R, Extraction kit, Alkaline lysis, Direct PCR

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DOI: https://doi.org/10.24002/biota.v6i2.3444

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