Ac-mon00110 Southern blot verification
Ac-mon00110
Southern blot verification:
Lane 1 = Ac-mon00110.1
Lane 2 = Ac-mon00110.2
Lane 3 = Ac-bti97156
Lane 4 = P-vv
Lane D = DIG VII ladder
Restriction Enzyme: EcoR1
Expected band size:. ~2.8 kb
Southern verification notes: Genomic DNA isolated from lines carrying Ac-mon00110 (lanes 1 & 2) or the progenitor line (lane 3), was digested with EcoR1, fractionated on agarose gels and transferred to nylon membranes. A DIG-labelled flanking sequence probe of 296 bp was generated using the Ac-end primer and flanking sequence primer (see PCR verification). Because EcoR1 cuts within the Ac element, the EcoR1 fragment shifts in the lines containing Ac-mon00110 relative to the progenitor line. Detailed descriptions of the Southern blot verification can be found in Brutnell and Conrad, 2003, Methods in Molecular Biology 236:157-76.