Ac-bti00228 PCR verification
Ac-bti00228 PCR
verification gel:
Lane 1 =TB00-228.2
Lane 2 = P-vv
Lane 7 = 100 bp ladder
Lane 8 = positive control
Lane 9 = negative control
Lane 10 = TB00-228.2
Lane 11 = TB00-228.3
Lane 12 = TB00-228.5
Lane 13 = TB00-228.6
Lane 14 = TB00-228.7
Lane 15 = TB01-255.1
Lane 16 = TB01-255.2
Lane 17 = TB01-255.3
Lane 18 = TB01-255.4
Lane 19 = TB01-255.5
Lane 20 = TB01-255.6
Lane 21 = TB01-255.7
Lane 22 = TB01-255.8
Lane 23 = TB01-255.9
Lane 24 = TB01-255.10
Lane 25 = P-vv
Lane 26 = 100 bp ladder
PCR verification primers:
Ac (TBp44) : 5’-TCGATCGGGATAAAACTAACAAA-3’
flanking (228.F1): 5’-GGGGCCTCCAAATTCCACTTCTA-3’
Product size: 461 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, 10 to 14, 16, & 20 to 24) that contained
the newly transposed Ac element. An Ac-specific primer was used
with a gene-specific primer to amplify the fragment of 461 bp. The absence of
this Ac from other progenitor lines results in the absence of the PCR product (lane
2 & 25). Detailed descriptions of Ac verification can be found in
Brutnell and Conrad, 2003, Methods in Molecular Biology, 236:157-76.