Ac-bti00228 Southern blot verification

Ac-bti00228 Southern blot verification:
Lane 1 = TB00-228.5
Lane 2 = TB00-228.2
Lane 3 = Ac-bti97156
Lane 4 = W22
Lane D = DIG VII ladder

Restriction Enzyme: Pst1
Expected band size:. ~6.8 kb

Southern verification notes: Genomic DNA isolated from lines carrying Ac-bti00228 (lanes 1 & 2) or not carrying Ac-bti00228 (lanes 3 & 4), was digested with Pst1, fractionated on agarose gels and transferred to nylon membranes. A DIG-labeled flanking sequence probe of 461 bp was generated using the Ac-end primer and flanking sequence primer (see PCR verification). Because Pst1 cuts within the Ac element, the Pst1 fragment has shifted in the lines containing Ac-bti00228 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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