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Comparing the Effectiveness of Varying the Amount of Manual Buffer Washes to Use of the QIAcube™ When Performing the QIAGEN MinElute® Post-PCR Clean Up

NCJ Number
Date Published
June 2009
9 pages
This project is a continuation of a recently completed study that used the QIAGEN MinElute Post-PCR cleanup to purify and concentrate amplified DNA on eight of the most common commercially available STR kits.
In the completed study, the evaluation team used four washes, the maximum number of washes recommended by the manufacturer. In the current study, the samples were subjected to the post-PCR cleanup process in which the number of washes varied from one to four, so as to determine the ideal number of washes that would yield the maximum result. In one procedure, the entire amplified product was cleaned up and concentrated with MinElute; all of it was placed on the Genetic Analyzer. Based on time and use of reagents, the study determined that two manual washes produced the best results. The QIAcube, however, produced the most consistent results from one concentration to another, probably because the pipetting is more consistent when using the robot compared to the manual processing. In a second procedure, all of the amplified product was cleaned up and concentrated with MinElute, and different volumes were added to the Genetic Analyzer. The volume added to the Genetic Analyzer influenced the fold increase; however, based on these results, there is not much difference between adding 5 ml or 10 ml. In a third procedure, different volumes of the amplified product were cleaned up and concentrated using the MinElute, and all resulting product was placed on the Genetic analyzer. Upon examination of heterozygosity, it was determined that after cleanup there was more consistency in the heterozygosity than previously. Regarding training requirements, minimal training is required to perform manual cleanups or to use the QIAcube. Both processes are relatively easy to perform. Regarding health and safety issues, the report cautions that QIAcube uses ethanol, a flammable liquid. Tabular data and a photo of the product

Date Published: June 1, 2009