Performing multiple PCR and extension reactions in a single well (i.e., multiplexing) is a way to increase the throughput and to reduce the effective cost per genotype. In other words, assays for different SNPs can be combined in a single reaction to save time, reagents, and to preserve DNA samples. MALDI-TOF MS provides the high resolution, high accuracy, and wide mass range needed to perform highly multiplexed genotyping assays.
To design SNP assays for multiplexing, several considerations must be taken. Among those:
1. Primers for the PCR amplification reactions must be designed in a way to avoid cross-loci amplification. In addition, PCR primer designs should have the following properties: 1) optimal primer length (20 mer); 2) optimal Tm (60oC); 3) optimal G-C content (50%); and 4) optimal amplicon length (~ 100 bp). Potential primers should be blasted against public databases to avoid individual assay failures due to competing kinetics or cross-hybridization.
5 ng human genomic DNA ~ 1600 copies
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The use of dumbbells gives you a much more comprehensive strengthening effect because the workout engages your stabilizer muscles, in addition to the muscle you may be pin-pointing. Without all of the belts and artificial stabilizers of a machine, you also engage your core muscles, which are your body's natural stabilizers.