CAJ | 학술논문

建立了测量噬菌体λ基因组DNA含量的方法。样品添加同位素标记碱基内标之后，用体积分数为88%的甲酸溶液在170℃水解30 min，解离出的核酸碱基通过反向柱分离，电喷雾四级杆质谱法测定，用多反应监测模式分别检测碱基及其同位素标记物的母离子和碎片子离子，从而建立了基因组DNA水解-同位素稀释质谱法测量长片段核酸含量的方法，并将DNA浓度溯源至碱基浓度。方法的线性范围为1～1000μg/g，检出限可低至100 ng/g。测定的λDNA含量标准物质为(2.51±0.06)μg/支（k=2），该方法可用于长片段核酸含量标准物质定值。
건립료측량서균체λ기인조DNA함량적방법。양품첨가동위소표기감기내표지후，용체적분수위88%적갑산용액재170℃수해30 min，해리출적핵산감기통과반향주분리，전분무사급간질보법측정，용다반응감측모식분별검측감기급기동위소표기물적모리자화쇄편자리자，종이건립료기인조DNA수해-동위소희석질보법측량장편단핵산함량적방법，병장DNA농도소원지감기농도。방법적선성범위위1～1000μg/g，검출한가저지100 ng/g。측정적λDNA함량표준물질위(2.51±0.06)μg/지（k=2），해방법가용우장편단핵산함량표준물질정치。
The method for determining genomic DNA mass content in phage was set up. Phageλgenome DNA samples were added by isotope labeled nucleic bases internal standard, then hydrolyzed in 88%formic acid solution for 30 min under 170℃. The DNA hydrolysis products nucleic bases were separated in C18 reverse phase column and analyzed by electrospray quadrupore isotope dilution mass spectrometry (HPLC-IDMS) by multiple reaction monitoring mode to scan parent ion and product ion of nucleic bases and the isotope labeled nucleic bases. The method was established for traceability of the genomic DNA to 4 nucleic bases mass content. The method linear range was 1-1 000μg/g, and the detection limit could be as low as 100 ng/g. The DNA mass content in each vial was measured (2.51±0.06)μg (k=2). This method could be used as a primary method for determining DNA mass as a reference material.