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

NCJ Number
136029
Journal
Journal of Forensic Sciences Volume: 37 Issue: 2 Dated: (March 1992) Pages: 597-603
Author(s)
A M Giusti; B Budowle
Date Published
1992
Length
7 pages
Annotation
This study describes the storage conditions for DNA bound to positively charged nylon membranes that impact positively and negatively on the ability to obtain restriction-fragment-length-polymorphism (RFLP) results.
Abstract
Human DNA from whole blood samples was isolated and analyzed according to procedures described by Budowle and Baechtel. Immediately after the DNA was fixed on the nylon membranes by heat baking at 80 degrees C for 30 minutes, the membranes were analyzed by hybridization with VNTR-specific probes or stored under various conditions. The membranes were stored under -20 degrees C, 4 degrees C, and ambient temperature and humidity. After varying times, the membranes were rehybridized to examine the effect of the various storage conditions. The concern was that storage conditions for the membranes permit RFLP profiles for the immobilized DNA over an extended period of time. Based upon study results, two brands of positively charged membranes examined can be stored at -20 degrees C or at 4 degrees C without any significant loss of detection sensitivity. Storage should be in sealed plastic bags or by thorough wrapping in plastic wrap at -20 degrees C to reduce the possibility of bacterial or fungal growth which might affect the integrity of the membrane or human DNA. 2 figures and 8 references

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