Ac-mon03080 PCR verification
Ac-mon03080 PCR
verification gel:
Lane 1 =100 bp ladder
Lane 2 = positive control
Lane 3 = TB01-346.1
Lane 4 = TB01-346.2
Lane 5 = P-vv
Lane 6 = W22
PCR verification primers:
Ac: 5’-TTTCCCATCCTACTTTCATCCCTG-3’
flanking (TB01-346.1F): 5’-CCTTCCTTCGCCTGTTTTTAGTGC-3’
Product size: 280 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 280 bp. The absence of
this Ac from other progenitor lines 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.