Illumina truseq and nextera mate pair library construction and sequencing

DNA was isolated from the spleens of five MutaMouse males (designated MutaMouse 1 through 5) using the QiaQuick Blood and Tissue kit (Qiagen). TruSeq paired-end libraries were prepared for all animals by shearing 1 µg of DNA with a Covaris ultrasonic disruptor followed by ligation of Illumina adapter sequences as per the manufacturer’s protocol. DNA from MutaMouse animal 2 was additionally used to prepare a Nextera Mate Pair library. Libraries were prepared by the Génome Québec Innovation Centre (Montréal, QC).

Low-coverage sequencing of MutaMouse 1 through 5 was carried out by the Génome Québec Innovation Centre (Montreal, Canada) using one lane of a HiSeq. 2500 (PE150). An entire additional HiSeq lane was used for the Nextera Mate Pair library of MutaMouse animal 2. The sequencing depth for all animals was increased by re-sequencing the same libraries in-house on a NextSeq. 500 (using the 300V2 sequencing kit) using one flow cell per animal sequenced. The two additional mice (male and female) from the Covance MutaMouse colony were used to construct Nextera Mate Pair libraries, which were then sequenced on the NextSeq. 500 as described above. All Illumina sequence reads produced were paired end, 150 bp. Data output from the sequencing are shown in Table 1.