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Mu transposon flanking sequences are amplified using the AFLP-like procedure detailed below. The enrichment of MluI linked fragments prior to PCR enables efficient and reproducible amplification of unmethylated Mu flanking sequences.

1. Digest genomic DNA with MluI and MseI
2. Ligate Mlu-Biotin adapter and MseI adapter, capture with streptavidin beads.
3. PCR amplify the Mu flanking sequences using a Mu specific primer and an MseI adapter specific primer.

Automated detection of amplified Mu flanking sequences.By substituting a fluorescent dye labelled primer during MuAFLP PCR, the number and size of the amplified Mu flanking sequences may be quantitatively determined on an automated genotyper such as an ABI 377 or MegaBACE. This provides a profile of Mu elements within individual plants which may be used to monitor the loss of insertions during backcrossing, or identify insertions co-segregating with a phenotype.

Mu flanking sequences were amplified from individual maize plants by MuAFLP using fluorescent labelled oligonucleotide. Fragments were fractionated on an ABI 377 and quantified using Genescan and Genographer software.

 
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Maintained by Gary Barker Last updated Dec 2001