Effect of Storage Conditions on Restriction Fragment Length Polymorphism (RFLP) Analysis of Deoxyribonucleic Acid (DNA) Bound to Positively Charged Nylon Membranes

    Volume 37, Issue 2 (March 1992)

    ISSN: 0022-1198

    CODEN: JFSOAD

    Page Count: 7


    Giusti, AM
    Physical scientist and research chemist, Forensic Science Research and Training Center, FBI Academy, Quantico, VA

    Budowle, B
    Physical scientist and research chemist, Forensic Science Research and Training Center, FBI Academy, Quantico, VA

    (Received 19 July 1991; accepted 2 August 1991)

    Abstract

    The ability to generate an autoradiogram from deoxyribonucleic acid (DNA) immobilized on a positively charged nylon membrane could be compromised by the storage conditions of the membrane. HaeIII-digested human DNA was size fractionated and transferred to two types of positively charged nylon membranes. The membranes were stored at −20°C, 4°C, and ambient temperature and humidity for times ranging from 1 day to 13 weeks, then hybridized to variable number of tandem repeat (VNTR) probes to examine the effect of the storage conditions on the membrane-bound DNAs. It was shown that such membranes could be successfully hybridized and rehybridized if they were stored at −20 or 4°C, but storage under ambient conditions reduced or eliminated the likelihood of successful hybridization.


    Paper ID: JFS13267J

    DOI: 10.1520/JFS13267J

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    Title Effect of Storage Conditions on Restriction Fragment Length Polymorphism (RFLP) Analysis of Deoxyribonucleic Acid (DNA) Bound to Positively Charged Nylon Membranes
    Symposium , 0000-00-00
    Committee E30