Ac-bti99224 Southern blot verification

Ac-bti99224 Southern blot verification:
Lane 1 = TB99-224.2
Lane 2 = TB99-224.1
Lane 3 = P-vv
Lane 4 = W22
Lane D = DIG VII ladder

Restriction Enzyme: EcoR1
Expected band size:. ~8.6 kb

Southern verification notes: Genomic DNA isolated from lines carrying Ac-bti99224 (lanes 1 & 2) or the progenitor lines (lanes 3 & 4), was digested with EcoR1, fractionated on agarose gels and transferred to nylon membranes. A DIG-labeled flanking sequence probe of 213 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 is larger in the lines containing Ac-bti99224 relative to the progenitor lines. Detailed descriptions of the Southern blot verification can be found in Brutnell and Conrad, 2003, Methods in Molecular Biology 236:157-76.


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