RT Journal Article SR Electronic T1 Inexpensive Multiplexed Library Preparation for Megabase-Sized Genomes JF bioRxiv FD Cold Spring Harbor Laboratory SP 013771 DO 10.1101/013771 A1 Michael Baym A1 Sergey Kryazhimskiy A1 Tami D. Lieberman A1 Hattie Chung A1 Michael M. Desai A1 Roy Kishony YR 2015 UL http://biorxiv.org/content/early/2015/01/21/013771.abstract AB Whole-genome sequencing has become an indispensible tool of modern biology. However, the cost of sample preparation relative to the cost of sequencing remains high, especially for small genomes where the former is dominant. Here we present a protocol for the rapid and inexpensive preparation of hundreds of multiplexed genomic libraries for Illumina sequencing. By carrying out the Nextera tagmentation reaction in small volumes, replacing costly reagents with cheaper equivalents, and omitting unnecessary steps, we achieve a cost of library preparation of $8 per sample, approximately 6 times cheaper than the widely-used Nextera XT protocol. Furthermore, our procedure takes less than 5 hours for 96 samples and uses nanograms of genomic DNA. Many hundreds of samples can then be pooled on the same HiSeq lane via custom barcodes. Our method is especially useful for re-sequencing of large numbers of full microbial or viral genomes, including those from evolution experiments, genetic screens, and environmental samples.