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KMID : 1101320080400020113
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Korean Journal of Clinical Laboratory Science
2008 Volume.40 No. 2 p.113 ~ p.117
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The Study of Nucleic Acid Extraction Method from Archival Paraffin Blocks
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Joo Kyung-Woong
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Abstract
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It designed a study to examine the efficiency of DNA and RNA extraction from archival formalin-fixed, paraffin-embedded tissues using an non-heating and heating method. Archival paraffin blocks of liver, kidney, colon were randomly selected. Each paraffin block was prepared in 20 microtubes. For each paraffin blocks were tested non-heating DNA extraction to 10 microtubes and heating protocol under pH 7.0 and 10 0℃ to 10 microtubes. Evaluation of the results of DNA extraction was carried out by measuring concentration by UV spectrophotometry and then PCR amplification. DNA extraction content that non-heating method was liver 5±0.7 μg/mL, kidney 2±0.3 μg/mL, colon 6±0.4 μg/mL and heating method was liver 12±0.6 μg/mL, kidney 7±0.5 μg/mL, colon 10.±0.3 μg/mL. Successful RNA extraction was observed, by β -actin amplification, in 46.7% sections for samples treated by the heating method versus 30.0% using non-heating DNA extraction. The extracted nucleic acid showed better values for samples heated at 100℃. Therefore heating extraction of nucleic acid is reliable, quick and efficiency.
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KEYWORD
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Paraffin-embedded tissue, Nucleic acid extraction, PCR, RT-PCR
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