Ac-mon00110 PCR verification
Ac-mon00110 PCR
verification gel:
Lane 1 = 1 kb ladder
Lane 2 = positive control
Lane 3 = negative control
Lane 4 = Ac-bti97156
Lane 6 = negative control
Lane 7 = Ac-bti97156
Lane 8 = Ac-mon00110.4
Lane 9 = positive control
Lane 10 = negative control
PCR verification primers:
Ac (900 3'end): 5’-CGTTACCGACCGTTTTCATCCCTA-3’
flanking (monki00-110R): 5’-AGCTGTTACGTGGAAGGTGCATAC-3’
Product size: 296 bp
PCR verification notes:
The DNA flanking the newly transposed Ac was cloned and sequenced from
one individual line. Verification PCR was performed using a line
that contained the newly transposed Ac
element (lane 8). An Ac-specific primer was used with a gene-specific primer to
amplify the fragment of 296 bp. The absence of this Ac from the progenitor
line results in the absence of the PCR product (lanes 4 & 7). Detailed descriptions
of Ac verification can be found in Brutnell and Conrad, 2003, Methods
in Molecular Biology, 236:157-76.