Figure 1.
Experimental strategy for recovering genome-wide insertion data. (A) Donor plasmid structure showing ssb tag in 3′ LTR. (B) The 25-bp tag is transferred to the 5′ end during retrotransposition, placing the tag within 24 bp of the 5′ end of the element. (C) A custom sequencing primer was designed to produce sequences consisting of 8 bp of Ty1 terminus and 30 bp of flanking sequences. (D) Genomic Southern blot to estimate copy number of new insertions. The first 12 lanes represent random colonies isolated from 12 independent transposition libraries. (p) Diluted donor plasmid lane.
