Ac-bti99221 PCR verification
Ac-bti99221 PCR
verification gel:
Lane 1 = positive control
Lane 3 = TB2309.1 (94)
Lane 4 = TB99-221.2
Lane 5 = W22
Lane 6 = P28
Lane 7 = negative control
Lane 8 = 100 bp ladder
PCR verification primers:
Ac (2901.9): 5’-ATCCCGTATTATCCCGTTCGTT-3’
flanking (P75.F1): 5’-CAGCCAGGGTTGTTTCTTCTTC-3’
Product size: 508 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 3 &
4) that contained the newly transposed Ac element. An Ac-specific primer
was used with a gene-specific primer to amplify the fragment of 508 bp. The
absence of this Ac from the additional lines, including a progenitor line, results in the absence
of the PCR product (lanes 5 & 6). Detailed descriptions of Ac
verification can be found in Brutnell and Conrad, 2003, Methods in Molecular
Biology, 236:157-76.