Optimization of In-house PCR Assay for Human BCL-2 Gene

    Henny Saraswati ,Anita Nauli Pulungan ,Aroem Naroeni ,Esa Fauzi
    Keywords: Apoptotic mechanism, BCL-2 gene, Polymerase Chain Reaction ,


    BCL-2 gene is present on human chromosome 18 and contributes in regulating the mechanism of
    apoptotic cells. Mutations and overexpression of the BCL-2 gene are known to be associated with several
    cancers. Comprehensive research on the role of BCL-2 in the incidence of cancer is needed to
    understand course of the diseases and treatment. Polymerase Chain Reaction (PCR) method for copying
    the BCL-2 gene can be used in this research. Optimization of the in-house PCR reaction was carried out
    to determine the optimal conditions of the PCR reaction on the doubling of the BCL-2 gene. In-silico
    analysis of the primer design produces three pairs of primers used in the optimization process. The
    parameters used in this optimization are the annealing temperature and primer concentration used.
    Validation of the results of optimization of the PCR reaction was carried out by biopsy samples of tumor
    tissue of patients with cervical tissue abnormalities. The result showed that the best primer pairs of the
    reaction is primer C with optimal annealing temperature is 570C and 800 nM concentration.

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