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In addition, the bioinformatic tools that can be used for sequence capture probe design as well as downstream sequence analyses are also addressed.
vaseyana used for sequence capture.
The probe set used for sequence capture contained 54773 probes representing 14729 P. taeda unigenes and are available in a previous study (Neves et al. 2013).
Thus, 13% of the sequence capture validated SNP positions also appeared to be heterogeneous (two nucleotides) within the collected individuals used for sequence capture.
From previous work (Neves et al. 2013) we expected that probes used for sequence capture could be capturing nonunique regions in the pine genome (e.g., paralogs and pseudogenes), hampering the detection of site-specific polymorphisms.
Each DNA extraction yielded greater than 2 μg of high molecular weight genomic DNA, which was used for sequence capture and Illumina HiSeq 2500 whole genome sequencing at the Institute for Genome Sciences, Baltimore, Maryland.
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The Nimblegen 385 K custom array was used for all capture protocols while the 454 GLX sequencer was used for sequencing.
Two TaMET1 ESTs as well as two brachypodium MET1 genomic fragments were selected and used to design probes for sequence capture (see Methods).
The nucleotide coding sequences of these genes were used to design probes for sequence capture, and a local sequence database was created with these sequences (subsequently called target database, TDB).
PCR primers were used for sequencing.
The extracts were further used for sequencing.
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