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ROUND 1. ROUND 2. 113,612 high-quality 3’-end ESTs. 147,146 high-quality 3’-end ESTs. CAP3 assembly. CAP3 assembly. 32,980 transcripts. 46,745 transcripts. Select PUTs with FLcDNA library clones Discard clones selected in round1. Select PUTs with FLcDNA library clones.
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ROUND 1 ROUND 2 113,612 high-quality 3’-end ESTs 147,146 high-quality 3’-end ESTs • CAP3 assembly • CAP3 assembly 32,980 transcripts 46,745 transcripts • Select PUTs with FLcDNA library clones • Discard clones selected in round1 • Select PUTs with FLcDNA library clones 9,759 candidate FL transcripts 10,710 candidate FL transcripts • Filter for FL candidate criteria • 2,771 missing 5’ SSPA • 1,312 short 5’- and/or 3’-end ESTs (Q20 < 400 nt) • 920 missing polyA tail • 357 5’-end ESTs > 100 nt shorter than public match • Filter for FL candidate criteria • 3,673 missing 5’ SSPA • 1,514 short 5’- and/or 3’-end ESTs (Q20 < 400 nt) • 1,119 missing polyA tail • 450 5’-end ESTs > 100 nt shorter than public match 2,779 candidates selected 5,348 candidates selected • Rearray into 96-well plates • Gap-closing by primer walking • Rearray into 384-well plates • 2x 3’-end and 2x 5’-end per clone • Manual and automated inspection for finished quality • Manual and automated inspection for finished quality 5,298 clones finished (65.1%) 1,166 clones finished (14.3%) 1,396 clones incomplete (17.1%) 267 clones aborted (3.2%)