Ac-bti00191 PCR verification
Ac-bti00191 PCR
verification gel:
Lane 1 = TB00-191.10
Lane 2 = P-vv
Lane 3 = 100 bp ladder
Lane 4 = TB00-191.1
Lane 5 = TB00-191.3
Lane 6 = TB00-191.4
Lane 7 = TB00-191.6
Lane 8 = TB01-227.1
Lane 9 = TB01-227.2
Lane 10 = TB01-227.3
Lane 11 = TB01-227.4
Lane 12 = 100 bp ladder
Lane 13 = positive control
Lane 14 = negative control
PCR verification primers:
Ac (232.9Ac): 5’-TGAAAATGAAAACGGTAGAGGTAT-3’
flanking (00-191.1F): 5’-CCACTCTGAATGAGAGTCACGAC-3’
Product size: 504 bp
PCR verification notes:
The DNA flanking the newly transposed Ac was cloned and sequenced from
one individual line. Verification PCR was performed using lines (lanes 1, 4 to 9, & 11) that contained the newly transposed Ac element. An Ac-specific
primer was used with a gene-specific primer to amplify the fragment of 504 bp.
The absence of this Ac from the progenitor line results in the absence
of the PCR product (lane 2). Detailed descriptions of Ac verification
can be found in Brutnell and Conrad, 2003, Methods in Molecular Biology, 236:157-76.