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Table 1 Comparison of standard versus improved solution hybrid selection methods

From: A scalable, fully automated process for construction of sequence-ready human exome targeted capture libraries

  Manual standard SHS protocol Automated improved SHS protocol
Process step Standard method Drawbacks Improved method Advantages
Shearing of genomic DNA Covaris S2 Single sample Optimized Covaris E210 Multi-sample, improved yield, tight size range
Enzymatic cleanups Individual spin columns Low throughput, 50 to 60% recovery, manual 'With-bead' SPRI High throughput, 80 to 90% recovery, automated
Solution hybrid selection capture Manual, column- based Labor intensive (6 samples/FTE/week) Fully automated Walkaway, high throughput (1,200 samples/4FTE/week)
Final PCR enrichment Denature, followed by PCR Sample loss through transfers Direct 'off-bead' PCR Improved final yield
In process quality control checkpoints Agilent Bioanalyzer Limited visibility until sequence results Many In process results: key predictors of sample, library and sequencing quality
  1. FTE, full time employee; SHS, solution hybrid selection; SPRI, solid phase reversible immobilization.