Straight walk: a modified method of ligation-mediated genome walking for plant species with large genomes.
Polymerase chain reaction (PCR)-based genome walking techniques are commonly used to clone unknown genomic regions flanking known sequences. However, these methods are typically problematic when applied to highly complex DNA templates isolated from plants with large genomes. Here we describe a reliable and efficient genome walking method that is particularly effective for plants with large genomes. Our ligation-mediated PCR method, Straight Walk, has improved sensitivity and specificity due to optimization of sequences of adaptors and adaptor primers. Successful genome walking in lily, which has one of the largest genomes in plants, indicates that Straight Walk is applicable for most plant species.
Tsuchiya
Tokuji
T
Graduate School of Science and Technology, Chiba University, 648 Matsudo, Chiba 271-8510, Japan.
Kameya
Nanako
N
Nakamura
Ikuo
I
D001483:Base Sequence; D017931:DNA Primers / Q000737:chemistry; D018745:Genome, Plant; D027762:Lilium / Q000235:genetics; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D017422:Sequence Analysis, DNA
D017931:DNA Primers