Extended Data Fig. 6: PCR confirmation of the site-specific T-DNA insertions identified by genome resequencing.
From: A mini foxtail millet with an Arabidopsis-like life cycle as a C4 model system
a and b. An Integrative Genomics Viewer (IGV) display of genome sequencing reads from WT (a) or the transgenic line N2 (b) spanning the T-DNA insertion site 22812363 on chromosome 7. The break point caused by the insertion is marked by an arrow. c. PCR confirmation of the insertion site 33288299 on chromosome 6 in line H2. d. PCR confirmation of the insertion site 22812363 on chromosome 7 in line N2. e. PCR confirmation of the insertion site 39094661 on chromosome 5 in line N8. Note: The genomic DNA for sequencing and PCR was prepared from pooling approximate 50 T1 transgenic seedlings, which explains the heterozygous nature of the T-DNA insertion seen in b-e. M, molecular marker; lane 1, no-transformed xiaomi plants; line 2, transgenic xiaomi plants; line 3, water control; F and R are primers for priming genomic regions flanking LB and RB ends of T-DNA, respectively; both the LB1 and LB2 primers are for T-DNA sequence close to the left border (LB). LB1 is 161 bp further apart from the border than LB2 for the vector pCAMBIA1305GFP, resulting in a band of bigger size in the R/LB1 pair in c. Similarly, LB1 and LB2 are distanced by 183 bp for the p8-GFP vector, thus resulting in different band size between F/LB1 and F/LB2 in d and e. All experiments were performed for three repeats, and similar results were obtained. Primers used are listed in Supplementary Table 3.
