Method for isolating RNA with high yield
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United States of America Patent
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Mar 23, 2021
Grant Date -
Jul 13, 2017
app pub date -
Jul 17, 2015
filing date -
Jul 17, 2014
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Abstract
A phenol-free method for isolating a nucleic acid from a sample is provided, said method comprising the following steps: a) preparing a precipitation mixture by adding at least one metal cation precipitant and at least one organic solvent selected from aprotic polar solvents and protic solvents to the sample, wherein the precipitation mixture i) comprises the metal cation precipitant; ii) comprises the organic solvent in a concentration of 15% or less; iii) comprises a buffering agent; and iv) has an acidic pH value, and precipitating proteins; b) separating the precipitate from the supernatant, wherein the supernatant comprises small RNA having a length of less than 200 nt and large RNA having a length of at least 1000 nt; and c) isolating a nucleic acid from the supernatant. Using an organic solvent as claimed during the protein precipitation step in the defined concentration provides a supernatant which in addition to small RNA also comprises large RNA. This is an advantage as the present method provides more flexibility to the user. The described method can be used for isolating and hence analysing different target RNAs.
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