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Arabidopsis 2010 Project:

WiscDsLox T-DNA Lines

 

Overview

Important Notice

DS Transposon

CRE/Lox Deletions

Find Insertions

Order Seeds

PCR primers

Vector Sequence

Seed Details

Questions?

AC Lines

Credits

 

 

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