Assay of deoxy-deazapurines in DNA by 3'-phosphorylation and two-dimensional thin-layer chromatography
作者： J.J. Steinberg , Gary W. Oliver , Antonio Cajigas
DOI: 10.1016/0378-4347(93)80174-3
关键词:
摘要: Abstract Deoxy-deazapurines (deaza-dNMPs) are incorporated into cellular DNA after administration of anti-neoplastic, anti-viral, or anti-parasitic chemotherapy. Deaza-dNMPs stable purine analogues and can be detected via 32P-labeling cold DNA. Assay analogue incorporation normal base composition is carried out by radiolabeling with all four deoxynucleotides (dNMPs) through nick translation. 3'-Monophosphate digest radiolabels representative dNMPs deaza-dNMPs. Separation occurs in two-dimensional polyethyleneimine—cellulose thin-layer chromatography, which resolves dNMPs. The technique was applied to human placental calf thymus DNA, control altered stoichiometric replacement deaza-dNMPs include deoxy-deazaadenosine, deoxy-deazaguanosine, deoxy-deazainosine. Scintillation detection densitometry both accurately reflect dNMP content. This easily quickly quantifies the low-molecular-mass deaza-dNMP Deaza-dNMP uptake may clinical chemotherapeutic efficacy host toxicity. assay therefore serve as an early biochemical dosimeter drug effect resistance.
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