Question: Can I add an extra shearing step during the Genome Protocol?
Answer: It is not recommended to add a shearing step to the Genome Protocol. During the Genome workflow, random priming of the oligo primer via the hexamer and a hexamer oligo supplied in the Master Mix, produces barcoded fragments ranging from a few to several hundred base pairs. The GEMs are then “broken”, pooling isothermally amplified, barcoded DNA molecules from all the different GEMs. Size selection is then used to optimize the insert size prior to library construction
Covaris shearing is not necessary as the SPRI cleanup optimizes the length of the average insert size. Covaris shearing will decrease library complexity and may cause increased PCR dup rates. This is similar to the result of adding an additional SPRI clean-up step.
Products: Genome, Exome