Ac-bti00245 PCR verification
Ac-bti00245 PCR
verification gel:
Lane 1 = TB01-264.2
Lane 2 = TB01-264.2
Lane 3 = TB01-264.3
Lane 4 = TB01-264.4
Lane 5 = TB01-264.8
Lane 6 = TB01-264.9
Lane 7 = TB01-264.9
Lane 8 = TB01-264.10
Lane 9 = postive control
Lane 10 = negative control
Lane 11 = 100 bp ladder
PCR verification primers:
Ac (TBp44): 5’-AAATCGGTTATACGATAACGG-3’
flanking (mon000245F.1): 5’-GATGCAAGCCTCACAGTCCACCTC-3’
Product size: 443 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 and
7) that contained the newly transposed Ac element. An Ac-specific primer
was used with a gene-specific primer to amplify the fragment of 443 bp. The
absence of this Ac from the additional sib line results in the absence
of the PCR product (lanes 2 to 6, & 8). Detailed descriptions of Ac
verification can be found in Brutnell and Conrad, 2003, Methods in Molecular
Biology, 236:157-76.