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FOUR STEPS USING BAC CLONES TO CONFIRM TGENIC EVENTS:
Step 1 Make a Custom BAC Library from the Plant Breeding Stack leaf tissue and pick/pool a strategic number of BAC clones.
Step 2 AEX optimizes probes and builds screening tools at the same time your Breeding Stack BAC library is being made.
Step 3 Using DNA/DNA hybridization, AEX scientists identify long gDNA BAC clones containing your Event(s) of Interest.
Step 4 AEX confirms the location of BAC clone ends & sequences the Event directly to verify Tgenic locus presence & position.
ADVANTAGES OF UTILIZING AMPLICON EXPRESS:
• Some Tgene targets can be difficult or impossible to amplify by PCR (AEX strategy does not rely on PCR for Event confirmation).
• BAC clones can simplify highly repetitive complex genomes down to only ~100Kb to 200Kb fragments.
• BAC clones allow you to determine haplotype & phase of each chromosome/allele.
• AEX lab is compatible with GLP requirements for regulatory approval.
• AEX has optimized Genome Reduction protocols to minimize # of clones screened.
• AEX can also use Fosmid cloning systems when needed (due to project constraints or HMW gDNA availability).
• Multiple Events can be screened simultaneously in a fast and cost effective manner.
• BAC clones serve as a permanent repository of your Tgenic Events (ideal for government agency approval).
• Amplicon Express (AEX) is world renowned for its excellence in BAC library construction and have made over 2,600 BAC libraries since 1996!
• AEX protocols are optimized for constructing high quality BAC libraries with fast turnaround times.
• This means your Breeding Stacks can be screened & Tgenic Events sequence-confirmed within a few weeks!
Details about Genome Reduction Strategy (to minimize cost):