METHOD FOR ISOLATING NUCLEIC ACIDS FROM FORMALIN-FIXED PARAFFIN EMBEDDED TISSUE SAMPLES

摘要

Methods are disclosed for isolating nucleic acids from formalin-fixed paraffin embedded (FFPE) tissue samples. Each of tissue samples contains paraffin and a target biological tissue or material, and the method includes the steps of: adding a first reagent and a second reagent to the FFPE tissue sample, the first reagent dissolving the paraffin material and the second reagent lysing the biological tissue; mixing the first reagent, the second reagent, and the FFPE tissue sample to form a first mixture; (2) heating the first mixture at 50-80° C. for 30-90 minutes; and then heating the first mixture at 80-95° C. for 30-90 minutes to fractionize the first mixture to form an aqueous phase and an oil phase; (3) collecting an aqueous solution from the aqueous phase; and (4) isolating nucleic acids from the aqueous solution. The method improves the efficiency and convenience of isolating nucleic acids from FFPE tissue samples.

主权项

1. A method for isolating nucleic acids from a formalin-fixed paraffin embedded tissue sample containing a paraffin material and a biological tissue, comprising the steps of:
(1) adding a first reagent, being an oily reagent and a second reagent, being an aqueous reagent to the formalin-fixed paraffin embedded tissue sample, the first reagent dissolving the paraffin material, releasing the biological tissue in the second reagent and the second reagent lysing the biological tissue, and releasing a nucleic acids, a protein containing in the biological tissue to the second reagent; mixing the first reagent, the second reagent, and the formalin-fixed paraffin embedded tissue sample to form a first mixture, wherein the oily reagent having a permittivity between 0 and 5, and containing straight-chain or branched-chain hydrocarbons having 7-10 carbon atoms with or without benzene groups, and the aqueous reagent lysing the protein in the biological tissue; (2) heating the first mixture at 50-80° C. for 30-90 minutes; and then heating the first mixture at 80-95° C. for 30-90 minutes to fractionize the first mixture to form an aqueous phase and an oil phase, wherein the nucleic acids dissolving in the aqueous phase; (3) collecting an aqueous solution from the aqueous phase; and (4) isolating the nucleic acids from the aqueous solution.