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Jurnal Ilmu Ternak dan VeterinerJurnal Ilmu Ternak dan Veteriner

The conventional polymerase chain reaction (PCR) method has become a prerequisite in molecular biology research. The PCR reaction is easy to prepare and only requires a small portion of the complex target nucleotide sequence, making PCR an easy and accurate method to use in biochemical and molecular analysis. PCR is generally divided into two categories: qualitative PCR and quantitative real-time PCR (RT-qPCR). The RT-qPCR method is more precise but has the disadvantage that it is much more expensive and requires more complicated equipment than conventional PCR. PCR products were visualized using agarose gel electrophoresis, which produced bands. Along with the development of digital technology, the resulting bands can be analyzed using digital software commonly used to analyze photos, such as ImageJ from the NIH. The trial results using ImageJ software to analyze CD44 compared to the housekeeping gene β-Actin demonstrated that gene expression can be quantified. Quantitative CD44 and β-Actin expression measurements were obtained by comparing the percentage of their respective peak plots. Analysis showed that CD44 expression was higher than β-Actin when evaluated with ImageJ software. These findings align with RT-qPCR results, which require more advanced PCR equipment and reagents. The semi-quantitative PCR analysis method using ImageJ offers a practical alternative for livestock and veterinary laboratories with limited budgets and resources.

The trial results using ImageJ software to analyze CD44 compared to the housekeeping gene β-Actin demonstrated that gene expression can be quantified.Analysis showed that CD44 expression was higher than β-Actin when evaluated with ImageJ software, aligning with RT-qPCR results.The semi-quantitative PCR analysis method using ImageJ offers a practical alternative for livestock and veterinary laboratories with limited budgets and resources.

Penelitian lebih lanjut dapat dilakukan untuk menguji akurasi metode analisis digital ini dengan berbagai jenis sampel dan penanda molekuler pada berbagai spesies ternak. Selain itu, pengembangan protokol standar dan antarmuka pengguna yang lebih ramah untuk perangkat lunak ImageJ akan mempermudah adopsi metode ini oleh laboratorium veteriner dan peternakan dengan sumber daya terbatas. Studi komparatif yang lebih mendalam antara hasil analisis ImageJ dengan metode kuantitatif PCR yang lebih mahal juga diperlukan untuk menentukan batas deteksi dan akurasi metode ini, serta mengidentifikasi potensi bias yang mungkin timbul.

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