Ac-mon00126 Southern blot verification:
Lane 1 = Ac-mon00126.4
Lane 2 = Ac-mon00126.5
Lane 3 = bti97156
Lane 4 = W22
Lane D = DIG VII ladder
Ac-mon00126 Southern blot
verification
Ac-mon00126 Southern blot verification:
Lane 1 = Ac-mon00126.4
Lane 2 = Ac-mon00126.5
Lane 3 = bti97156
Lane 4 = W22
Lane D = DIG VII ladder
Restriction Enzyme: Pst1
Expected band size:. ~5.0 kb
Southern verification
notes: Genomic DNA isolated from lines carrying Ac-mon00126 (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 155 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 PstI
fragment is 4.5 kb larger in the lines containing Ac-mon00126 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.