Abstract:

BACKGROUND: Because of the damage of formalin-fixed paraffin-embedded tissue to DNA in the process of its production and preservation, it is difficult to extract high quality and sufficient DNA for PCR.
OBJECTIVE: To analyze the influencing factors of the DNA extracted from paraffin-embedded tissue for PCR.
METHODS: Genomic DNA was extracted from paraffin-embedded esophageal carcinoma tissue sections of 10.0 μm×2,      10.0 μm×4 and 10.0 μm×5. Template DNA amount of 0.05, 0.1, 0.2 μg was used to amplify gene β-actin, respectively. PCR cycle times were set as 35, 40 and 45. The influencing factors of PCR were analyzed.
RESULTS AND CONCLUSION: The analysis of agarose gel electrophoresis showed that when paraffin-embedded esophageal carcinoma tissue section amount was 10.0 μm×2, the PCR cycle times were 40; when the template DNA amount was 0.05μg, the obtained target DNA was of the highest quality. It shows that reducing the amount of paraffin-embedded tissue and DNA template can be help to require high quality PCR strips; PCR cycle times should be more than 40 and less than 45, and if it increases beyond this range there is meaningless.