Improvement of HER2I655V TARMS-PCR Performance by DNA Quality Analysis

Improvement of HER2I655V TARMS-PCR Performance by DNA Quality Analysis

Bugi Ratno Budiarto, Azamris, and Desriani

Annales Bogorienses Vol. 21 (2): 52-62.

 tarmReliable TARMS-PCR is a prerequisite in constructing a solid conclusion in genetic diagnostics. The validity of data generated by this molecular technique in most cases is hampered by a false positive result. In attempt to develop a TARMS-PCR for HER2I655V genotyping with no interfering of bias we used DNase I to eliminate DNA contaminant resided in PCR reagent. TARMS-PCR with no enzymatic pretreatment on PCR master mix kit produced a false positive result on HER2I655V TARMS-PCR using as recombinant plasmids system model proven by the presence of multiple PCR products in Non-Template Control (NTC). A dose of 0.1 U of the enzyme could eliminate this DNA contaminant effectively, although this pretreatment altered the specificity of HER2I655V TARMS-PCR genotyping on certain genotype. Combination of touchdown TARMS-PCR with another allelespecific primer recovered specificity of detection on this model system. Interestingly, this optimized HER2I655V TARMS-PCR can only be used for genotyping the clinical samples if only further optimization was done using genomic DNA as template.

Keywords: TARMS-PCR, HER2I655V, DNase I, Polymorphism

Katalog: http://

Disusun oleh: Ludya AB/Pustakawan


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