Ac-mon00126 PCR verification
Ac-mon00126
PCR verification gel:
Lane 1 = 1 kb ladder
Lane 2 = positive control
Lane 3 = negative control
Lane 4 = bti97156
Lane 5 = W22
Lane 6 = Ac-mon00126.4
Lane 7 = Ac-mon00126.5
PCR verification primers:
Ac (900 3'end): 5’-CGTTACCGACCGTTTTCATCCCTA -3’
flanking (mon000126F): 5’-AGTGAGCTAGCATGATGCACGAAC-3’
Product size: 155 bp
PCR verification notes:
The DNA flanking the newly transposed Ac was cloned and sequenced from
one individual line. Verification PCR was performed using the original line
(lane 6), and a sib (lane 7) that also contained the newly transposed Ac
element. An Ac-specific primer was used with a gene-specific primer to
amplify the fragment of 155 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.