BAC sequence was determined by NGS on an Illumina HiSeq 1000 using a 100 bp paired-end protocol or an Illumina MiSeq using a 300 bp paired-end protocol. The DNA library was prepared from seven BAC pools (details shown in Additional file 1 : Table S1, S8, S10, and S11) using TruSeq DNA Sample Prep v2 (Illumina), following the manufacturer’s instructions, with a unique identifier index added to each pool for multiplex sequencing.
Truseq dna sample prep v2
Manufactured by Illumina
The TruSeq DNA Sample Prep v2 is a laboratory equipment product designed for the preparation of DNA samples. It provides a standardized workflow for the processing and library preparation of DNA samples prior to sequencing.
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Lab products found in correlation
3 protocols using truseq dna sample prep v2
1
BAC Genome Sequencing Using NGS
2
Comprehensive Genome Sequencing Approaches
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WGS pyrosequencing (GS-FLX-Titanium) was performed according to the manufacturer protocols (Roche Diagnostics) yielding 8 kb paired-end reads. Illumina 2×100 paired-end WGS was performed on a GAIIx at Eurofins MWG Operon and/or locally using TruSeq DNA Sample Prep v2 and TruSeq SBS v5 kits (Illumina). These methods provided >99 % genome coverage at an average depth of >20× and >500×, respectively, for 52 genomes. In addition, paired-end Illumina data (2×250 cycles) were obtained using Nextera XT DNA Library Preparation and MiSeq Reagent V2 (500 cycles) kits on a MiSeq instrument according to the manufacturer's instructions (Illumina) yielding >100× coverage.
3
Whole-Genome Library Preparation and Sequencing
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We generated a whole-genome paired-end library with the Illumina TruSeq DNA Sample Prep v2 (Illumina, San Diego, CA), selecting for a mean fragment length of 450 bp with the SPRIselect reagent kit (Beckman Coulter, Krefeld, Germany). A mate-pair library with an insert size of 5 kb was created with the Nextera Mate Pair Sample Prep Kit (Illumina). The paired-end and mate-pair libraries were sequenced on an Illumina MiSeq machine. Long-read sequencing was performed on the PacBio RS II system (Pacific Biosystems of California, Menlo Park, CA), using 16 SMRT cells in total.
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