Ac-mon03077 PCR verification
Ac-mon03077 PCR
verification gel:
Lane 1 = P-vv
Lane 2 = Ac-mon03077
Lane 3 = 100 bp ladder
Lane 4 = mon00222.1
Lane 5 = mon00222.2
Lane 6 = mon00222.3
Lane 7 = mon00222.4
Lane 8 = mon00222.5
Lane 9 = 100 bp ladder
PCR verification primers:
Ac: 5’-CCCGCAAGTTAAATATGAAAATGA-3’
flanking: 5’-TACTCCGGGTGGTGCTCTGAATAA-3’
Product size: 820 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 2 & 4 to 8) that contained
the newly transposed Ac element. An Ac-specific primer was used
with a gene-specific primer to amplify the fragment of 820 bp. The absence of
this Ac from other progenitor lines results in the absence of the PCR product (lane
1). Detailed descriptions of Ac verification can be found in
Brutnell and Conrad, 2003, Methods in Molecular Biology, 236:157-76.