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The number of the reads matched barcode was 813497032 (94.6%).
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Matching barcodes were assigned to the SD control and heat stress sample (TGACCA), as well as the SC control and heat stress samples (GCCAAT), to allow for multiplexing on two lanes of an Illumina GA-II genome analyzer flow cell.
The barcode sequences, number of associated reads, and the experimental description matching each barcode are shown in Additional file 1: Table S1.
So the reads matched barcodes while not matched BsaXI recognition sites were analyzed.
With "best close match" the best-matching reference barcode is identified, but the query is only assigned the taxon name of that barcode if the barcode is sufficiently similar (i.e. below a threshold).
Using the Greengenes database, the Bacteroidetes DNA barcode was less specific, demonstrating sequence matches with a greater number of phyla (7 of 15).
A "false-positive" result, where a query barcode is matched to a reference barcode despite significant divergence, is a frequent consequence of using the BLAST algorithm by itself [ 33].
During our analysis it was found that a proportion of reads had incomplete barcodes which could nevertheless be rescued and so only those with a perfect match to the M13 primer and at least four bases of the barcode (and which could be reconstructed back to a unique, full barcode) were used.
Determine if a unique barcode is needed.
Samples were processed in batches of 24 to match barcodes available.
The matching of the correlation patterns to the reference barcodes was carried out by visual comparison between the investigative and reference spectra.
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