Evaluation of DNA extraction from archieved formalin-fixed paraffin tissues (FFPT)‎ by B-globin gene PCR

الملخص EN

Background : the knowledge of the nucleic acid state before start to molecular detection have a great importance to make a math measurements by which the nucleic acid quantity and quality will be knows.

Objective : Conventional methods have limitations for extracting amplifiable DNA from specimens containing a small number of cells.

DNA quality and quantity was evaluated by β-globin amplification and NanoDrop instrument respectively.

DNA extraction from FFPT has become essential for molecular research specially with advances in molecular technology.

Methods : The current study tested breast cancer FFPT specimens of Iraqi women which collected from AL-Sader teaching hospital histopathology and neoplasm units, and several privet histopathology labs in AL-Najaf province, the specimens were stored for the years (2008, 2009, 2010 and 2011).

The specimens were included of BC 54, fibroadenoma 21, mastitis 18, and ductectesia 19 samples.

Results : Specific primers were used to detect the activity of beta-globin gene by PCR technique and the results appeared that The mean of DNA quantity was (61.68 ng / ul) while the DNA extract quality which appeared positive by β-glubine PCR was 71.42 % (80 of 112), BC 70.37 % (38 of 54), fibroadenoma 85.71 % (18 of 21), mastitis 77.77 % (14 of 18) and ductectesia 52.63 % (10 of 19), there was a significant difference (p < 0.5).

The beta-globin PCR appeared different results depending on the DNA stats.

Conclusions : the evaluation of the extracted DNA by NanoDrop system and beta-globin gene is the easiest and definite method compared with other researchers who worked of other evaluation methods.

Recommendations : This study recommends that every study performed on DNA should be started with beta-globin testing to avoid false negative results caused by degraded DNA.