RESUMO
A two-step chemical method of introduction of nonradioactive labels in DNA was proposed. At first step DNA is modified by succinic dihydrazide at pH 5.0 and 95 degrees C, or at pH 4.5 and 37 degrees C in presence of sodium bisulfite. Then FITC or biotin are joined to the hydrazide groups. DNA modified in this way were shown to be effective hybridisation probes.
Assuntos
Sondas de DNA , DNA Viral/efeitos dos fármacos , Hidrazinas/farmacologia , Succinatos/farmacologia , Bacteriófago lambda/genética , Biotina , Fenômenos Químicos , Química , DNA Viral/genética , DNA Viral/metabolismo , Hidrazinas/metabolismo , Indicadores e Reagentes , Hibridização de Ácido Nucleico , Succinatos/metabolismo , Sulfitos , Fagos T/genéticaRESUMO
A two-step chemical method has been developed for the introduction of biotin and other low-molecular derivatives into DNA. The method is based on the interaction of aliphatic diamines with cytosine residues of the polynucleotide chain in the presence of sodium bisulfite with subsequent addition of biotin. DNA modified this way may be used as an effective probe for non-isotopic hybridization which can be used simultaneously with 32P-labelled probes.