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Optimizing Large-Scale EST Assembly: Efficient Selection Protocol for Unigenes

This non-duplicate EST assembly protocol involves selecting and assembling unigenes using Phrap, removing undesirable sequences, and rearraying ESTs for efficient processing. The steps include probe delivery, bioinformatics analysis, and validation to remove duplicates.

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Optimizing Large-Scale EST Assembly: Efficient Selection Protocol for Unigenes

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  1. Non-duplicate Unigenes Selection Protocol Large-scale EST assembly using phrap Contigs Unassembled ESTs Unigenes Remove undesirable sequences, such as, retrotranspsons, organellar gene sequences, E. coli and phage DNA sequences and rRNAs Rearray selected ESTs to 384-well plates Plasmid DNA preps PCR amplification of inserts 5’ sequencing 3’ sequencing Sequence processing EST validation and duplicates removal Probe delivery Bioinformatics Lab Bioinformatics Lab

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