Ac-mon00200 PCR verification
Ac-mon00200 PCR
verification gel:
Lane 1 = 100 bp ladder
Lane 2 = W22
Lane 3 = Ac-bti97156
Lane 4 = Ac-mon00200.4
Lane 5 = Ac-mon00200.5
PCR verification primers:
Ac (900 3’end): 5’-CGTTACCGACCGTTTTCATCCCTA -3’
flanking (mon00200F.1): 5’-TGGAACTTCTACGCAGCAGCA -3’
Product size: 289 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 289 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.