Effect of storage conditions on restriction fragment length polymorphism (RFLP) analysis of deoxyribonucleic acid (DNA) bound to positively charged nylon membranes.
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 degrees C, 4 degrees 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 degrees C, but storage under ambient conditions reduced or eliminated the likelihood of successful hybridization.
Forensic Science Research and Training Center, FBI Academy, Quantico, VA.
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AM
Giusti
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B
Budowle
D001345:Autoradiography; D004247:DNA / Q000032:analysis*; D015342:DNA Probes; D006801:Humans; D006813:Humidity; D009693:Nucleic Acid Hybridization; D009757:Nylons; D012150:Polymorphism, Restriction Fragment Length; D011309:Preservation, Biological; D013696:Temperature
D015342:DNA Probes; D009757:Nylons; D004247:DNA