Ac-mon00152 PCR verification
Ac-mon00152 PCR
verification gel:
Lane 1 = 100 bp ladder
Lane 2 = Ac-bti97156
Lane 3 = W22
Lane 4 = Ac-mon00152.5 (plasmid)
Lane 5 = Ac-mon00152.5 (genomic)
Lane 6 = negative control
Lane 7 = positive control
PCR verification primers:
Ac (900 3'end): 5’-CGTTACCGACCGTTTTCATCCCTA-3’
flanking (monki000152F.1): 5’-TAGGGCTAGGCGTTGTGCAG-3’
Product size: 196 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 4 & 5) that contained the newly transposed Ac element. An Ac-specific
primer was used with a gene-specific primer to amplify the fragment of 196 bp.
The absence of this Ac from the progenitor line results in the absence
of the PCR product of the appropriate size (lanes 2 & 3). Detailed descriptions of Ac verification
can be found in Brutnell and Conrad, 2003, Methods in Molecular Biology, 236:157-76.