1,N6-乙烯基脱氧腺苷和3,N4-乙烯基脱氧胞苷的高效液相色谱-32P后标记分析
HPLC-32P-postlabelling analysis of 1,N6-ethenodeoxyadenosine and 3,N4-ethenodeoxycytidine.
作者信息
Watson W P, Crane A E
机构信息
Shell Research Ltd, Sittingbourne Research Centre, Kent, UK.
出版信息
Mutagenesis. 1989 Jan;4(1):75-7. doi: 10.1093/mutage/4.1.75.
A 32P-postlabelling procedure coupled with HPLC has been developed to detect and measure the cyclic nucleic acid adducts 1,N6-ethenodeoxyadenosine and 3,N4-ethenodeoxycytidine in DNA. Chloroacetaldehyde-modified DNA containing these adducts was enzymatically digested to 3'-monophosphates and adducts were separated by ion-pair reverse-phase HPLC on PL RP-S prior to 32P-postlabelling with carrier free [gamma-32P]ATP. Following 3'-dephosphorylation with nuclease P1 the resulting [5'-32P]monophosphate adducts were finally resolved by HPLC on PL RP-S and assayed by liquid scintillation counting.
已开发出一种结合高效液相色谱（HPLC）的³²P后标记程序，用于检测和测量DNA中的环状核酸加合物1,N⁶-乙烯基脱氧腺苷和3,N⁴-乙烯基脱氧胞苷。含有这些加合物的氯乙醛修饰DNA经酶消化为3'-单磷酸酯，加合物在无载体[γ-³²P]ATP进行³²P后标记之前，通过离子对反相HPLC在PL RP-S上进行分离。用核酸酶P1进行3'-去磷酸化后，所得的[5'-³²P]单磷酸加合物最终通过在PL RP-S上的HPLC进行分离，并通过液体闪烁计数进行测定。
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