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In that case, barcoding power could be increased by using both barcoded forward and reverse PCR primers, but this would restrict the length of the sequenced regions, as each region would only be sequenced in one direction.
Clearly large crop collections cannot be sequenced in one draft.
In an appropriate experimental design, thousands of samples can be sequenced in one run.
Such an index read allows for multiple samples to be sequenced in one lane (multiplexing).
Second, the unique barcoding of each recombinant/mutant allowed hundreds to be sequenced in one or a few SMRT cells, which greatly reduces the overall sequencing cost.
However, since Illumina sequencing is highly sensitive to templates presented in DNA samples, some traced transcripts or contaminants can be sequenced in one sample but not in other samples.
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Libraries prepared from the 95 samples and one blank negative control were sequenced in one lane on an Illumina HiSeq 2500 (Illumina, San Diego, California, USA).
Three biological replicate RNA samples from each stage of the leaves were sequenced in one lane on the Illumina Hi-Seq 2500, and the number of reads ranged from 5,000,000 to 18,000,000.
Then the library pool was sequenced in one GS FLX Titanium run on 1/8th of a 70 × 75 PicoTiter plate.
Pooled DNA from parents, 189 RILs, 94 other lines and 3 blanks was sequenced in one lane with the Illumina HiSeq sequencer at Genomic Diversity Facility, Cornell University Institute of Biotechnology (http://www.biotech.cornell.edu/brc/genomic-diversity-facility).
Each sample was sequenced in one lane of an Illumina Genome Analyzer IIx sequencer.
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