Ac-mon00068 PCR verification
Ac-mon00068 PCR
verification gel:
Lane 1 = Ac-mon00068.6
Lane 2 = Ac-mon00068.7
Lane 3 = Ac-mon00068.8
Lane 4 = W22
Lane 5 = P-vv
Lane 6 = negative control
Lane 7 = positive control
Lane 8 = 100 bp ladder
PCR verification primers:
Ac (900 3'end): 5’-CGTTACCGACCGTTTTCATCCCTA -3’
flanking (mon00068F2): 5’-CTACTGACTGCCATTGATTTCTTC -3’
Product size: 183 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 1,2 & 3) that contained the newly transposed Ac element. An Ac-specific
primer was used with a gene-specific primer to amplify the fragment of 183 bp.
The absence of this Ac from the progenitor line results in the absence
of the PCR product (lanes 4& 5). Detailed descriptions of Ac verification
can be found in Brutnell and Conrad, 2003, Methods in Molecular Biology, 236:157-76.