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Selection and Finalization of FLcDNA Candidates from High-Quality ESTs for Poplar Gene Cloning

This study focuses on the systematic selection and refinement of full-length cDNA (FLcDNA) candidates from a pool of over 90,000 high-quality 3'-end ESTs, identifying 5,926 potential candidates. Through stringent filtering for cloning structures and sequencing quality, including the removal of entries lacking key components such as 5' SSPA and polyA tails, we narrowed down our list to 5,443 finalized FL candidates, alongside 4,664 finished sequences. The methodology employed ensures high accuracy and reliability for subsequent cloning efforts.

eugenia-norris
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Selection and Finalization of FLcDNA Candidates from High-Quality ESTs for Poplar Gene Cloning

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  1. B A Candidate selection FLcDNA finishing 90,368 high-quality 3’-end ESTs 5,926 FL candidates • Filter for cloning structure • missing 5’ SSPA (stringent) • missing polyA tail (stringent) CAP3 assembly 35,011 putative unique transcripts (PUTs) 5,653 FL candidates • Filter for rearray accuracy • mixed well • rearray to correct plate PUTs lacking clones derived from FL libraries excluded 7,346 FL candidates 5,532 FL candidates • Filter for FL candidate criteria • short 5’- and/or 3’-end ESTs (Q20 < 100 nt) • shorter at 5’ end than public poplar ESTs • missing 5’ SSPA • missing polyA tail • Filter for sequencing problems • hard stop 5,525 FL candidates • Filter for clone features • sequence vs. gel data • chimeric 5,926 FL candidates 5,443 FL candidates + 4,664 (79%) finished 779 (13%) incomplete 483 (8%) aborted

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