Abstract

MARQUEZ, Caterin et al. Comparison of four methods of DNA restoration in samples from plasma and pap smears as a tool for better the samples. Rev. colomb. cienc. quim. farm. [online]. 2011, vol.40, n.2, pp.189-200. ISSN 0034-7418.

Objetives: To compare four methods of restoration of DNA in plasma and PAP smears as a tool to improve the quality of the samples. Methods: 20 blood samples and 20 PAP smears samples, we performed DNA isolation by commercial kit and phenol-chloroform respectively. Then all samples underwent a pre-PCR treatment with four different types of activity DNA polymerase: 1. Exonuclease and endonuclease 5'-3'. 2. Exonuclease 5'-3'. 3. Klenow, and 4. Klenow more ligase. Different restoration methods were evaluated quantitatively by real-time PCR with gene ALU. Results: All restoration methods improve the quality of DNA in both types of samples. However, the 3th method showed better results in both plasma and PAP smears, increasing the concentration of DNA from 0.0022 ng/mL to 0.6474 ng/mL in PAP smears and 0.0039 ng/mL to 0.435 ng/mL in blood plasma. Conclusions: DNA from plasma samples and PAP smears to be treated with a restoration process increases the quality of DNA compared to untreated samples.

Keywords : PAP smears; plasma; restoration; polymerase chain reaction; Taq polymerase.

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