Ac-mon00236 PCR verification
Ac-mon00236 PCR
verification gel:
Lane 1 = Ac-bti97156
Lane 2 = W22
Lane 3 = Ac-mon00236.6
Lane 4 = Ac-mon00236.8
Lane 6 = positive control
Lane 7 = 1 kb ladder
PCR verification primers:
Ac (700 5’ end): 5’-TTTCCCATCCTACTTTCATCCCTG -3’
flanking (mon00236F.1): 5’-CGCCGTACCTAGCACGTTTT -3’
Product size: 229 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 3 &
4) that contained the newly transposed Ac element. An Ac-specific
primer was used with a gene-specific primer to amplify the fragment of 229 bp.
The absence of this Ac from the progenitor line results in the absence
of the PCR product (lanes 1 & 2). Detailed descriptions of Ac verification
can be found in Brutnell and Conrad, 2003, Methods in Molecular Biology, 236:157-76.