Ac-mon00030 PCR verification
Ac-mon00030 PCR
verification gel:
Lane 1 = 100 bp ladder
Lane 2 = positive control
Lane 3 = negative control
Lane 4 = Ac-bti97156
Lane 5 = W22
Lane 6 = Ac-mon00030.3
Lane 7 = Ac-mon00030.4
Lane 8 = 100 bp ladder
PCR verification primers:
Ac (900 3'end): 5’-CGTTACCGACCGTTTTCATCCCTA-3’
flanking (mon00030F): 5’-TATTAGTCAACGGGTCGTGTCGTG-3’
Product size: 261 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 6 & 7) that contained the newly transposed Ac element. An Ac-specific
primer was used with a gene-specific primer to amplify the fragment of 261 bp.
The absence of this Ac from the progenitor line results in the absence
of the PCR product (lanes 4 & 5). Detailed descriptions of Ac verification
can be found in Brutnell and Conrad, 2003, Methods in Molecular Biology, 236:157-76.