Ac-bti00209 PCR verification
Ac-bti00209 PCR
verification gel:
Lane 1 = 100 bp ladder
Lane 2 = TB00-209.1
Lane 3 = TB00-209.7
Lane 4 = TB00209.11
Lane 5 = TB00-209.12
Lane 6 = TB01-241.1
Lane 7 = TB01-241.2
Lane 8 = TB01-241.3
Lane 9 = TB01-241.4
Lane 10 = TB01-241.5
Lane 11 = P-vv
Lane 12 = 100 bp ladder
PCR verification primers:
Ac (242.7Ac): 5’-GATAACGGTCGGTACGGGATTTTC-3’
flanking (mon000209.2F): 5’-GTAGAAGAAGACGCGCTCCACG-3’
Product size: 340 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 to 10) that contained
the newly transposed Ac element. An Ac-specific primer was used
with a gene-specific primer to amplify the fragment of 340 bp. The absence of
this Ac from the progenitor line results in the absence of the PCR product
(lane 11). Detailed descriptions of Ac verification can be found in Brutnell
and Conrad, 2003, Methods in Molecular Biology, 236:157-76.