Accurate Normalization of Grain Samples Using Pressure-Based Volume Measurement Technology
Poster Feb 07, 2015
Bill Gigante, John Thomas Bradshaw, Tanya Knaide
A common challenge experienced by sample screening facilities is identifying individual sample quantity prior to DNA extraction. Accurately identifying sample quantity allows the user to pipette exact amounts of extraction buffer into each well. Herein, we present the normalization of a unique sample type, namely kernels of a variety of grains. The variability of kernel size was determined in each well of a 96-well microplate with subsequent normalization to a total volume of 200 µL. Using initial volume measurements collected with a new pressure-based volume measurement technology, the required volume of buffer was accurately determined for each grain variety in every well of the microplate. This new technology is embodied in an instrument, the Artel VMS, which uses air pressure to measure the volume contained within individual wells independent of plate type, sample material, or well shape to rapidly and accurately identify individual volumes in both 96-well and 384-well microplate formats.
We utilized paired synthetic crRNAs coupled with our synthetic tracrRNA in cells transduced with lentiviral Cas9 to perform a functional knockout on hsa-miR-221. This three-part system (crRNA, tracrRNA and Cas9) has demonstrated efficient gene editing when used with only one guide RNA, but the goal was to use two crRNAs to remove the entire stem-loop.READ MORE