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Fresh leaves were dried overnight at 55°C and DNA was extracted using the extraction kit Dneasy® from Qiagen®.
For neoplasia-free control group, DNA was obtained from peripheral blood white cells using the extraction kit Mini Spin (Biometrix, Curitiba, PR, Brazil), according to manufacturer's instructions.
Total RNA was extracted from mucosa samples by the method of Chomczynski and Sacchi (1987) [ 21] using the extraction kit from Bangalore Genei, India.
PCR products were purified from agarose gels using the extraction kit Wizard SVGel and PCR clean-up system (Promega) and then inserted into pCR2.1-TOPO vector to chemically transform competent E. coli cells (One-Shot TOP10, Invitrogen).
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A single colony from each plate was removed for DNA extraction using the RBC Genomic DNA extraction kit (RBC Bioscience, Chung Ho City, Taiwan) as per the manufacturer's directions.
Peripheral blood samples collected in EDTA tubes were subjected to DNA extraction using the GE extraction kit (GE Healthcare Lifesciences BR).
The second sample was subjected to DNA extraction using the DNeasy extraction kit (Qiagen, Hilden, Germany) according to the manufacturer's instructions.
Size selection (200 400 bp) of the final PCR product was done by agarose gel electrophoresis and gel extraction using the Nucleospin extraction kit (Macherey & Nagel).
The MDA method performed excellent using the ENA extraction kit and poorly when using the QIA extraction kit.
Bacterial genomic DNA extraction was done using the ZR Soil Microbes DNA Mini-Prep extraction kit (Zymo Research Corporation, South Africa).
Total RNA was extracted using the "RNeasy extraction kit" from Quiagen (Hilden, Germany).
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