Ac-mon00068 Southern blot verification

Ac-mon00068 Southern blot verification:
Lane 1 = Ac-mon00068.6
Lane 2 = Ac-mon00068.7
Lane 3 = P-vv
Lane 4 = W22
Lane D = DIG VII ladder

Restriction Enzyme: Pst1
Expected band size:. ~5.1 kb

Southern verification notes: Genomic DNA isolated from lines carrying Ac-mon00068 (lanes 1 & 2) or the progenitor lines (lanes 3 & 4), was digested with Pst1, fractionated on agarose gels and transferred to nylon membranes. A DIG-labeled flanking sequence probe of 183 bp was generated using the Ac-end primer and flanking sequence primer (see PCR verification). Because Pst1 does not cut within the Ac element, the Pst1 fragment is shifted in the lines containing Ac-mon00068 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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