Illumina HiSeq 1500 paired end sequencing; Draft genome sequencing of Lactuca. sativa cv. Tizian
Draft genome sequencing of Lactuca. sativa cv. Tizian
After two months of cultivation, a young plant at the six leaf stage was selected, flash frozen and ground to a fine powder using a sterile pestle and mortar with liquid nitrogen. Lettuce ( Lactuca sativa var. longifolia) cv. Tizian was cultivated under controlled 'axenic' conditions. Prior to cultivation, the industrial seed coating was removed with a sterilized pestle and mortar. The seeds were washed with sterile water and subsequently surface sterilized in 5% sodium hypochlorite for 15 minutes. Afterwards, the seeds were washed again three times with sterile water. The seeds were placed on Petri dishes containing a Murashige and Skoog Basal Salt Mixture (MS) agarose medium (4.4 g per liter, Sigma-Aldrich Chemie GmbH) and incubated in a cultivation chamber with a 16/8 h day/night cycle and a temperature of 16 °C. After germination, the seedlings were transferred to Petri dishes with a MS (4.4 g per liter) agarose medium supplemented with sucrose (30 g per liter) in order to check for microbial contaminations.  DNA was extracted with the DNeasy Plant Mini Kit (Qiagen, Hilden, Germany). Two sequencing libraries were constructed, one Nextera Mate-Pair library with a Mate-pair distance length of 9 kb on average and a TruSeq PCR-free whole genome shotgun library (Illumina Inc., San Diego, U.S.A.).
Experimental Factor: organism
Lactuca sativa var. longifolia