GENE PANELS
METHODOLOGY
In essence there are 2 mainstream methods to enrich for specific genomic targets : (1) the hybrid capture method and (2) the PCR based approach.
The hybrid capture based method is more lenient towards 'content updates' (addition or removal of probes/targets), but is more laborious.
The PCR based approach generally has a better limit of detection and is less laborious, however it requires a longer or more complex optimization phase in case multiple targets need to be enriched for.
1. Hybrid capture method
Fragmentation of genomic DNA
End repair of fragmented DNA + phosphorylation and adenylation
PCR amplification of the probe-selected fragments
Fragmentation of genomic DNA
2. PCR based method
Amplification of target of interest with locus specific tagged primers
Nested PCR on the tagged PCR amplicons (step 1) in order to incorporate a sample specific barcode (grey) and flowcell recognition sites (red and green)
Amplification of target of interest with locus specific tagged primers
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