Skip to main content

Advertisement

Figure 1 | BMC Genomics

Figure 1

From: A Bead-based Normalization for Uniform Sequencing depth (BeNUS) protocol for multi-samples sequencing exemplified by HLA-B

Figure 1

Schematic workflow of BeNUS. The BeNUS method was constructed using two categories of method, size selection (A) and normalization of DNA amount (B). (A) Size selection using altered AMPure XP beads. For size selection, two different bead ratio conditions were applied according to DNA volume: 0.4× bead ratio for <1,000 bp fragment size and 0.5× bead ratio for >500 bp. After bead selection, DNA fragments ranging from 500 to 1,000 bp were bound to the beads. (B) Normalization of DNA amount using altered AMPure XP beads DNA fragments with target sizes ranging from 500 to 1,000 bp were selected for effective HLA gene haplotype phasing. The size selection and DNA amount also defined an actual molar concentration for bridge PCR to generate clusters in a flow cell, because DNA fragments of over 1,000 bp are not efficiently amplified. Only one bead reaction condition was applied to normalize the amount of DNA. This step enables a defined amount of DNA to be bound to the diluted beads. This step enables a defined amount of DNA to be bound to the diluted beads.

Back to article page