An improved protocol to increase sensitivity of Southern blot using dig- labelled DNA probes

Montserrat Solanas, Eduard Escrich

    Research output: Contribution to journalArticleResearchpeer-review

    6 Citations (Scopus)

    Abstract

    The use of methods for nonradioactive labelling of nucleic acids has increased in recent years because they avoid disadvantages associated with radioisotopes. The most frequently used label is digoxigenin (DIG). The greatest problem of nonradioactive methods is their high nonspecific background mainly caused by the multistep detection. A diffuse background can mask the specific signal; furthermore nonspecific signals can make it difficult to interpret the result. In this study we have attempted to identify elements which could generate background. We have also determined the probe and antibody concentrations by which the higher sensitivity is obtained.
    Original languageEnglish
    Pages (from-to)153-159
    JournalJournal of Biochemical and Biophysical Methods
    Volume35
    DOIs
    Publication statusPublished - 3 Dec 1997

    Keywords

    • Chemiluminescence
    • Digoxigenin
    • Non-radioactive
    • Sensitivity
    • Southern Blot

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