ISSN: 0022-1198
CODEN: JFSCA
Page Count: 7

Effect of Storage Conditions on Restriction Fragment Length Polymorphism (RFLP) Analysis of Deoxyribonucleic Acid (DNA) Bound to Positively Charged Nylon Membranes
Giusti, AM
Physical scientist and research chemist, Forensic Science Research and Training Center, FBI Academy, VA

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

Keywords:
pathology and biology, deoxyribonucleic acid (DNA), hybridization, restriction fragment length polymorphisms, storage conditions, nylon membranes, variable number of tandem repeats

Paper ID: JFS13267J
DOI: 10.1520/JFS13267J
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