Ac-bti99224 PCR verification
Ac-bti99224 PCR
verification gel:
Lane 1 = TB99-224.1
Lane 2 = TB99-224.2
Lane 3 = W22
Lane 4 = P-vv
Lane 5 = positive control
Lane 6 = negative control
Lane 7 = 100 bp ladder
PCR verification primers:
Ac (P77Ac.1): 5’-GACCTGTACCGACCGTTTT-3’
flanking (P77.F1): 5’-TGATCGAAATCCAGTGAACG-3’
Product size: 213 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 & 2) that contained the newly transposed Ac element. An Ac-specific primer
was used with a gene-specific primer to amplify the fragment of 213 bp. The
absence of this Ac from the progenitor lines results in the absence
of the PCR product (lanes 3 & 4). Detailed descriptions of Ac
verification can be found in Brutnell and Conrad, 2003, Methods in Molecular
Biology, 236:157-76.