Equipment:
Roche Lightcycler 480-II
Details:
Quantitative PCR allows for highly accurate, and cost effective measurement of fluorescent-labeled nucleic acids. This system can be used to quantify cDNA isolated from mRNA and miRNA levels from cells or tissues. The instrument also allows quantitative discriminations of DNA for genotyping, haplotype and SNP analyses.
The LightCycler 480-II is a leading machine in the mid throughput range of instrumentation, boasting a silver thermal block with rapid heating (4.80C/sec) and cooling (2.50C/sec) rates. The machine’s Therma-base technology allows precise experimental homogeneity minimizing well variability throughout the block. The LightCycler 480 has 5 excitation filters (440, 465, 498, 533 and 618nm), 6 emission filters (488, 510, 580, 610, 640 and long pass filter: 660nm), and an internal barcode scanner. Fluorescent dyes compatible with the system include: Cyan 500, FAM/SYBR, VIC/HEX, LC Red 610, LC Red 640, Cy5, Yakima Yellow, Cal Orange, Tamara, Cy3, Alexa 546, Cal Red 610, ROX, Texas Red and Alexa 647. The LightCycler 480-II system includes software for complex analyses, including absolute quantification using the standard curve method, crossing point determination using the 2nd derivative max method, the proprietary “E-method” for quantitative assessments, and a genotype module for melt curve and endpoint calls.
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