Flow chart illustrating the physical map contig-specific fragment preparation.
figureposted on 2013-10-24, 03:22 authored by Yanliang Jiang, Parichart Ninwichian, Shikai Liu, Jiaren Zhang, Huseyin Kucuktas, Fanyue Sun, Ludmilla Kaltenboeck, Luyang Sun, Lisui Bao, Zhanjiang Liu
The minimal tilling path BAC clones from each physical contig were selected (highlighted). The pooled BAC DNA from each physical map contig was digested with two 4-bp restriction enzymes, Mse I and Bfa I, respectively. The digestion product was then ligated with in-house designed adaptors, followed by PCR using in-house designed primers. The combination of adaptor and primer formed a specific tag representing each physical contig ID. All PCR products with a physical map contig-specific tag then were pooled together, and sequenced using Illumina HiSeq 2000 platform.