While most nucleic acids are intracellular, trace amounts of DNA and RNA are present in peripheral blood. Many studies have suggested the potential utility of these circulating nucleic acids in prenatal diagnosis, early cancer detection, and infectious diseases diagnosis. However, DNA circulating in blood is usually present at very low concentrations (ng/ml), and is in the form of small fragments (<1,000 bp), making its isolation for diagnostic purposes challenging. Here we report an improved method for isolation of small DNA fragments from serum using the groove-binding, polycationic compaction agents to selectively precipitate DNA. A 151 bp Lambda DNA fragment served as the model DNA in our experiments.  Using compaction precipitation, we were able to recover and detect very low levels (0.01 ng/ml) of a small DNA fragment in serum. The isolated DNA product after compaction precipitation was largely free of serum contaminants and suitable for downstream applications, such as sequencing.

Recovery of small diagnostic DNA fragments from serum using compaction precipitation

243rd National Meeting (2012)

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