February 14, 2019 by Team Pillar
In NGS library prep using traditional multiplex PCR, paired primers flank individual regions of interest to generate PCR amplicons. When amplifying contiguous gene targets with overlapping regions, the innermost primers from each primer pair will generate amplicons, which, due to the nature of PCR, can quickly overwhelm the reaction mix. To overcome this issue of overlapping amplicons, traditional methods require separate PCR reactions that are then pooled into a single library.
Stem-Loop inhibition-mediated amplification (SLIMamp®) is a proprietary target enrichment chemistry that enables overlapping multiplex PCR for efficient target amplification in a single tube. With SLIMamp chemistry, tag sequences are added to primer pairs such that overlapping amplicons have complementary regions that anneal to form stem-loop structures that become poor templates for further amplification of these undesirable amplicons. SLIMamp chemistry gives you the ability to tile amplicons across any genomic region of interest all in a single reaction tube.
Click below to watch an animation that highlights the details of SLIMamp enrichment chemistry