Ac-mon00072 PCR verification
Ac-mon00072 PCR
verification gel:
Lane 1 = 100 bp ladder
Lane 2 = Ac-bti97156
Lane 3 = W22
Lane 4 = Ac-mon00072.1
Lane 5 = Ac-mon00072.3
Lane 6 = positive control
Lane 7 = negative control
PCR verification primers:
Ac (700 5' end): 5’-TTTCCCATCCTACTTTCATCCCTG-3’
flanking (mon00072F700): 5’-CTTCACCACGCACATAGAGAGAGA-3’
Product size: 225 bp
PCR verification notes:
The DNA flanking the newly transposed Ac was cloned and sequenced from
two pooled lines. 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 225 bp.
The absence of this Ac from the progenitor line results in the absence
of the PCR product (lanes 2 & 3). Detailed descriptions of Ac verification
can be found in Brutnell and Conrad, 2003, Methods in Molecular Biology, 236:157-76.