The Y Chromosome Deletion Detection System, Version 2.0, provides a standardized screening panel amplifying only informative nonpolymorphic sequence tag sites (STS) on the human Y chromosome. The system amplifies key functional regions associated with AZoospermia Factor (AZF), namely the regions that flank AZFa and cover AZFb, AZFc, AZFd including DAZ, KAL-Y, SMCY and flanking loci for other key spermatogenesis-related genes (namely RBM1, DFFRY and DBY). Five Multiplex Master Mixes, with a total of 20 characterized Y-specific primer pairs, are included. Four of the multiplex primer sets contain a control primer pair that amplifies a fragment of the X-linked SMCX locus. One of the multiplex primer sets (Multiplex E Master Mix) contains a control primer pair that amplifies a unique region in both male and female DNA (ZFX/ZFY). Finally, a primer pair that amplifies a region of the SRY gene has been included in Multiplex E Master Mix as a control for the testis-determining factor on the short arm of the Y chromosome to detect XX males arising from Y to X translocations. The Multiplex Master Mixes are designed to facilitate the simultaneous amplification of several different regions of the Y chromosome. The amplification products (83-496bp) of the five multiplex PCR amplifications can be clearly separated by agarose gel electrophoresis and visualized by ethidium bromide staining. Failure to amplify specific regions of the Y chromosome is indicative of Y chromosome deletions in the test sample. The size control ladder provided minimizes analysis time and the possibility of misinterpreting molecular weight of amplification products.|
Ease of Use: Premixed Multiplex Master Mixes contain 20 primer pairs, including internal controls providing a standardized panel of results requiring no user optimization. More Robust Reactions: Improved formulation and use of GoTaq(R) DNA Polymerase minimizes dropouts. Flexibility: Amplify genomic DNA purified using various methods and with a PE480 (oil overlay) or PE9600/9700 (non-oil overlay) thermal cycler. Complete System: All required reagents are provided in the kit.