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Assess genomic DNA size, quality, and quantity
Quality assessment of genomic DNA is essential in a number of downstream applications, including PCR, genotyping, microarray, and next-generation long-read sequencing. Evaluating the viability of gDNA samples in a time eﬃcient manner is essential prior to sequencing and other downstream applications.
The Fragment Analyzer™ Automated CE System streamlines gDNA analysis with the DNF-467 Genomic DNA 50 Kb Analysis Kit, allowing the operator to size, qualify, and quantify high concentration samples with a single 200-fold dilution, requiring an input concentration range of 25 ng/µL - 250 ng/µL.
An electropherogram overlay of genomic DNA samples is shown above with the corresponding digital gel image. Assessment of sample quality can be handled eﬃciently through the use of the Genomic Quality Number (GQN). The user designates a threshold in PROSize ® Data Analysis Software, which then assesses each sample as it relates to the threshold assigning a value between 0 and 10. Values are determined based on how much of the sample exceeds the threshold. In the example above, a GQN threshold of 10,000 bp was deﬁned (designated by the dotted blue line) with calculated values shown in the table above.Two-fold serial dilutions of gDNA were made and subsequently analyzed by the Fragment Analyzer and a ﬂuorometric instrument. The electropherogram and digital gel image depicting these dilutions are shown above along with a table showing the associated concentration values. Genomic DNA concentrations obtained by both instruments were compared and plotted on a graph. The results showed a one-to-one linear relationship between the ﬂuorometric instrument and the Fragment Analyzer. The accuracy of the Fragment Analyzer was <15% and precision was <20%.