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METHOD OF SEPARATING EXTRA-CHROMOSOMAL DNA FROM OTHER CELLULAR COMPONENTS
METHOD OF SEPARATING EXTRA-CHROMOSOMAL DNA FROM OTHER CELLULAR COMPONENTS
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机译:从其他细胞成分中分离染色体外DNA的方法
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摘要
THE INVENTION RELATES TO A METHOD OF SEPARATING EXTRA-CHROMOSOMAL DNA FROM RNA. IT ALSO RELATES TO DNA PRODUCED BY THE METHOD AND PHARMACEUTICALS DERIVED FROM SUCH DNA, FOR EXAMPLE, DNA VACCINES. THE METHOD COMPRISES SEPARATING EXTRA-CHROMOSOMAL DNA FROM RNA WITHOUT FIRST DIGESTING THE RNA, AND COMPRISES:@@i) LYSING CELLS COMPRISING EXTRACHROMOSOMAL DNA TO FROM A LYSATE;@ii) CLARIFYING THE LYSATE; AND @ii) SUBJECTING THE CLARIFIED LYSATE TO TANGENTIAL FLOW ULTRA FILTRATION UNDER CONDITIONS, WHICH ALLOW A SUBSTANTIAL AMOUNT OF THE RNA TO PASS THROUGH A MEMBRANE WHILST THE EXTRA-CHROMOSOMAL DNA IS RETAINED. THE PREFERRED CONDITIONS INCLUDE THE USE OF A DIAFILTRATION BUFFER OF LOW CONDUCTIVITY AND DIAFILTRATION VOLUMES OF OVER 10 VOLUME EQUIVALENTS.
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