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Abstract
Background and purpose: Assessment of genetic changes from a molecular viewpoint is essential for diagnosis, follow-up and treatment. For molecular procedures, DNA extraction is the first step. Therefore, applying economical and easy procedures will be helpful. Pathological specimens are normally fixed in formalin and paraffin after surgery and it is important to extract quality DNA from specimens. This study is intended to compare Chelex and Phenol / Chloroform methods of DNA extraction in Paraffin-embedded Colorectal Cancer specimens and their effect on PCR-SPSS.
Methods and Materials: This descriptive-analytical study was conducted on 60 colorectal cancer specimens including 10 rectal and 50 various colon specimens. DNA extractions were done by Chelex and Phelex / Chloroform methods and their quality and quantity were measured by spectrophotometer. The P53 gene was amplified using specific primers in a PCR assay and, consequently, electrophoresis was done. The two methods were compared and analyzed by Kappa coefficient.
Results: Out of 60 subjects, 15 were female and 45 male. Their age ranged from 44 to 91 with the mean being 62 years. Quality DNA was extracted from 44 formalin-fixed specimens by Chelex method, and the PCR was positive. However, with the same sample, 32 specimens had suitable PCR assay by Phenol / Chloroform method. Therefore, a significant relationship was observed between the two methods (p
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