Ac-mon00186 PCR verification
Ac-mon00186 PCR
verification gel:
Lane 1 = W22
Lane 2 = Ac-bti97156
Lane 3 = Ac-mon00186.4
Lane 4 = Ac-mon00186.5
Lane 9 = negative control
Lane 10 = positive control
Lane 11 = 100 bp ladder
PCR verification primers:
Ac (700 5’end): 5’-TTTTCCCATCCTACTTTCATCCCTG -3’
flanking (mon00186.32F.1): 5’-GGGCAGCAAACTCGTGATTC -3’
Product size: 236 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 236 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.