Orange 500 Fluorescent Size Standard User manual Version: 1.0 Revision date: 08-02-2019 Product and Company Information Product name: Product use: Company: Telephone: Email: Orange 500 Fluorescent Size Standard (p/n DSMO-100) For Research Use Only NimaGen BV Lagelandseweg 56 6545 CG Nijmegen The Netherlands +31 (0)24 820 02 41 [email protected] Description The Orange 500 Fluorescent Size Standard is a fluorescent labeled ultra-accurate size standard for the reproducible sizing of fragment analysis peaks on any ABI Genetic Analyzer with a five-dye spectral calibration. The standard contains 16 Orange dye-labeled, single-stranded fragments in the 45-595 nucleotides range with basepair lengths of 35, 50, 75, 100, 139, 150, 160, 200, 250, 300, 340, 350, 400, 450, 490 and 500 bp. The size standard can be used for a variety of applications like Microsatellites, MLPA, Fragment Length analysis, Fragment Length Polymorphisms etc. Each of the DNA fragments is labeled with a LIZ® compatible fluorophore resulting in a sharp orange peak under denaturing conditions. With this size standard, your own custom marker fragments can be labeled with the dyes FAM™, VIC® , NED™ or PET® (G5 filterset, Life Technologies) or with FAM™, ZEL™, SOL™, FER™ (B5 filterset, Biolegio/Nimagen) . Each kit contains enough standard for at least 800 analyses. Protocol Before use, mix the contents of each tube thoroughly and centrifuge briefly to collect the liquid at the bottom of the tube. Typical loading mix: 0.5 µl (diluted) sample, 0.5 µl Size Standard and 9 µl Super-DI or Hi-Di™ Formamide. This cocktail should be considered as a starting point only. Optimize these ratios if necessary, based on your experimental results. If needed, extend the time for datacollection in your instrument software, to obtain collection of the full length