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Kashif A. Haque, Casey L. Dagnall, Adri O’Neil, and Belynda D. Hicks, Core Genotyping Facility, Advanced Technology Program, SAIC-Frederick, Inc.,
The robustness of many SNP genotyping technologies forgives inaccurate and imprecise liquid transfers of DNA and reagents. However, multiple automated liquid transfers during typical DNA sample handling and genotyping workflows can lead to inaccurate and imprecise DNA estimates and reagent consumption. Variability can lead to inefficient consumption of precious DNA aliquots and reagents, and a reduction in actual DNA mass versus expected can lead to lower completion and higher discordance rates. The National Cancer Institute’s Core Genotyping Facility (CGF) has implemented QA/QC measures in all laboratories, analytical, and in silico workflows. A Laboratory Automation Maintenance Plan (LAMP) has been established to ensure the accuracy and precision of eight Beckman Coulter Biomek FX and NX laboratory automation workstations used in DNA sample handling and SNP genotyping platforms. The LAMP consists of volume calibration with fluorescein of the Multichannel and span-8 pods on all workstations (see Figure 1.), “run-only” access to automation methods for lab staff, and SOPs. Calibration is performed for all robotic tips (Multichannel) and syringe sizes (Span-8), for all desired transfer volume ranges. Robotic versus manual accuracy is plotted and the resulting “Scaling Factor” (slope) and “Offset” (y-intercept) are adjusted for each instrument’s liquid handling techniques. Following a series of internally-created SOPs, robotic accuracy (~ 5% vs. manual) and precision (%CV of < 5%) has been achieved for all desired transfer volumes (2 to 200 uL). All procedures are performed quarterly and data is stored electronically. Implementation has led to a measurable increase in completion and concordance rates in all genotyping platforms.. Click here for this poster (PPT)
Additional Information
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