PCR primers

Use the following PCR primer to confirm the presence of a desired T-DNA insertion in a line:

        p745:   5'-AACGTCCGCAATGTGTTATTAAGTTGTC-3'

The primer p745 hybridizes to the T-DNA Left Border region of pDs-Lox. 

The T-DNA Left Border sequence from pDS-Lox is shown below with the p745 primer sequence highlighted in RED:

5’-TAGCTCTTGCTAAGCTCCTCGAGTTTCTCCATAATAATGTGTGAG
TAGTTCCCAGATAAGGGAATTAGGGTTCCTATAGGGTTTCGCTCA
TGTGTTGAGCATATAAGAAACCCTTAGTATGTATTTGTATTTGTA
AAATACTTCTATCAATAAAATTTCTAATTCCTAAAACCAAAATCC
AGTACTAAAATCCAGATCCCCCGAATTAATTCGGCGTTAATTCAG
TACATTAAAAACGTCCGCAATGTGTTATTAAGTTGTCTAAGCGTC
AATTTGTTTACACCACAATATATCCTGCCA-3’
 

The T-DNA/Plant DNA junction typically occurs in the region highlighted in BLUE. 

The Arabidopsis genomic DNA that flanks a T-DNA insertion site will be found on the 3' side of the blue region shown above.

NOTE:  The primer that we used when we did the high-throughput DNA sequencing reactions for this project was p745.