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The 13,200 clones obtained from both libraries were then submitted to fosmid DNA extraction in pools and subsequent pyrosequencing.
Subsequently, all the CCS reads from both libraries were then aligned to determine a base-substitution error frequency at each position.
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Fosmid DNA extracted from all pools for both libraries was then combined to produce a single DNA sample, a master pool of 5 μg DNA, for subsequent pyrosequencing.
RNA from both libraries was then fragmented in a buffered zinc solution and single stranded RNA adapters were ligated to the 5′ and 3′ ends to maintain strand specificity, prior to re-purifying on a polyacrylamide gel.
Both genomic and transcriptomic libraries were then mapped to the identified clusters of genomic 454 data using BLAT (Kent 2002).
cDNA libraries were then sequenced using a HiSeq1000 Illumina system.
The selected or weighted libraries were then used to analyze the ELECTRA reactor.
The resulting Fab combinatorial libraries were then screened for binding to the antigen.
Libraries were then prepared and indexed using the DNA Ultra II Library Preparation Kit (New England Biolabs, E7645).
Amplified libraries were then purified using Agencourt AMPure XP beads and eluted in 30 μl EB.
Libraries were then quantified using the PicoGreen Life Technologiess) and pooled up to 12-plex.
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