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Initially, about 200 clones were partially sequenced from each library to assess library quality.
Ninety six clones were selected at random and sequenced to assess library diversity.
Sequences from each cDNA library were closely monitored to assess library complexity and sequence quality.
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.
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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).
To account for unigene redundancy per gene we assessed library-specific read frequencies per UL rather than per unigene.
The motivation behind our current work was to determine the lower limits of metagenomic library preparation protocols and to assess which library prep performed best down to single picogram DNA input levels.
There was no way you could assess this library, so Lou pulled a price out of his head, a lowish price, because it was just a nightmare.
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