CPC C12N 15/1017 (2013.01) [B01D 61/145 (2013.01); B01D 71/12 (2013.01); C12P 19/34 (2013.01); G01N 30/02 (2013.01); G01N 2030/027 (2013.01)] | 45 Claims |
1. A method for producing formulated purified RNA, comprising the steps of:
A1) providing a plasmid DNA encoding a RNA polymerase promoter sequence, a RNA coding sequence, and a restriction endonuclease cleavage site, wherein the coding sequence for said RNA is no greater than 15,000 nucleotides in length;
A2) linearizing the plasmid DNA with a restriction endonuclease to produce linearized DNA;
A3) ultrafiltering the linearized DNA by tangential flow filtration (TFF), to produce purified linearized DNA;
B1) transcribing the purified linearized DNA to produce transcribed RNA;
B2) treating the transcribed RNA with a DNase;
C1) ultrafiltering the transcribed RNA by performing at least one step of TFF using a TFF membrane cassette having a molecular weight cutoff (MWCO) of less than or equal to 500 kDa, to produce purified RNA; and
D) formulating the purified RNA to produce the formulated purified RNA.
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