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We quantified and assessed library quality on a Bioanalyzer 2100 and randomly pooled equimolar samples in four lanes of the Illumina flowcell (eight or nine samples per lane).
We quantified and assessed library quality on a Bioanalyzer 2100 and, within each species, randomly pooled equimolar samples onto two or three lanes of the Illumina flowcell (8 10 samples per lane).
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To account for unigene redundancy per gene we assessed library-specific read frequencies per UL rather than per unigene.
Initially, about 200 clones were partially sequenced from each library to assess library quality.
Sequences from each cDNA library were closely monitored to assess library complexity and sequence quality.
Ninety six clones were selected at random and sequenced to assess library diversity.
A portion of each reaction was analyzed by electrophoresis on a 1.5% agarose gel to assess library quality.
After trial PCR to assess library quality and quantity, 30 μl cDNA was run on a native 6% PAGE gel.
To assess library stability, 14 random BAC clones were assayed by serial culture for more than 100 generations over a period of 5 days.
To assess library complexity by determining the maximum number of unique reads obtainable from a MP library, two of the large-insert libraries (20-kb and 25-kb) were sequenced to a higher depth in an additional sequencing run.
We assessed the library quality by assaying ligations and carrying out 5′-end sequencing; the former procedure determined library titer, and the latter used to evaluate cDNA full-length percentage as well as the proportion of empty vectors.
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