High-purity DNA extraction from animal tissue using picking in the TRIzol-based method

Abstract

TRIzol is used for the extraction of RNA, DNA and proteins from tissues or cells. Here, we present a simple picking method to extract DNA from tissues using TRIzol. Spectrophotometric analysis showed that the 260/280 and 260/230 nm optical density ratio of the picking method's DNA is ideal and better than that obtained by the classic TRIzol method. Gel electrophoresis showed that there was no RNA contamination, and the DNA had not degraded. DNA extracted by the picking method had the same performance in restriction enzyme digestion and quantitative PCR as that obtained by the traditional method. Viral DNA in the infected tissue was also obtained. This modified method facilitates various molecular biology assays.

METHOD SUMMARY

In the traditional step of extracting DNA with TRIzol reagent, DNA flocculent clumps will be visible after adding absolute ethanol and mixing upside down, picking the DNA clumps with a pipette tip instead of through centrifugation. The picked DNA was transferred to 0.1 M sodium citrate in 10% ethanol and rinsed twice, then rinsed once in 75% ethanol. It was then dried and dissolved in 8 mM NaOH.