DNA甲基化检测技术解读.pptVIP

感谢聆听！ THANK YOU FOR WATCHING! 演示结束！ * Part 2 of our afternoon double feature is the instrumentation aspect of this amplicon melting application. We currently have 2 instruments designed specifically for Hi-Res melting, … the LightScanner and the HR-1. The LightScanner is available in a 96 or 384 well format. The heat block used is compatible with almost any currently available microtiter plate used on a variety of block style thermal cyclers. Melt time is approximately 5 minutes per plate which gives melting curve results on all of the samples in the plate. Just to be clear, 5 minutes of melting gives you 96 or 384 results, which is approximately 4 seconds per sample in a 96 well plate, and 1 second per sample in a 384 well plate. The heat block in the LightScanner is capable of maintaining a +/- 0.2 degree difference between wells across the plate, which is pretty good considering even the most accurate thermal block cyclers typically are capable of maintaining only a +/- 1 degree difference across the plate. At a thermal ramp rate of 0.15 degrees/second, the LightScanner collects 20 data points per degree. The HR-1 is a single sample instrument, designed around amplification in a LightCycler capillary. The HR-1 accepts the capillary in a custom designed heating element called an “ingot”. Basically, the ingot is made of a highly conductable metal that is wrapped in a heat coil such that there is no detectable temperature gradient from top to bottom or side to side in the capillary. This assures temperature homogeneity within the sample and a more accurate depiction of what is happening to the sample as it is melted. Scanning of a single sample takes approximately 30 seconds in the HR-1, and the optics are custom designed for the LCGreen dye. As I mentioned, the ingot design is capable of maintaining at least a +/- 0.05 degree intra- and inter-run temperature difference, and may actually be better than that, but 0.05 degrees is the limit of what we could acc