OPTIMIZATION OF PCR BASED ON DNA QUALITY IN ONCOLOGICAL RISK POPULATION DNA SAMPLES FROM BENIN
In Benin, like other countries in the sub-Saharan region, breast cancer ranks first among all cancers. Material and methods: A highly heterogenic collection of 120 DNA samples from breast and prostate cancer patients and matched controls was available for further study on detection of BRCA1/2 gene mutations pattern. Results: Initially, the PCR amplification of BRCA1 exon 5 was performed for all samples, with poor results in terms of PCR efficiency. Only about a third of the samples showed visible bands of the expected size and with no contamination. A re-evaluation of DNA samples showed high variability on DNA concentration and quality, due to different extraction methods, sample origin and preservation condition. Conclusions: Our study primarily illustrate that DNA extracted from different tissue by different extraction methods may be damaged but still usable, by increasing the DNA amount in the PCR reactions and adjusting the other PCR parameters. However, appropriate caution should be exercised when analyzing samples derived from different tissue sources. By appropriate optimization, heterogenous samples can be simultaneously used for PCR amplifications with single parameters.
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