1. Uso da bioinformática na análise genômica

2. TAGAGCATCGATCGATGCTGCAGATGATGCTAGCATCGGCTAGGCGACG ATCTCGTAGCTA ATCTCGTAGCTAGCTACGACGTCTA ATCTCGTAGCTAGCTA ATCTCGTAGCTAG ATCTCGTAGCTAGC ATCTCGTAGCTAGCT ATCTCGTAGCTAGCTAC ATCTCGTAGCTAGCTACG ATCTCGTAGCTAGCTACGA ATCTCGTAGCTAGCTACGAC ATCTCGTAGCTAGCTACGACG ATCTCGTAGCTAGCTACGACGT ATCTCGTAGCTAGCTACGACGTC ATCTCGTAGCTAGCTACGACGTCT ATCTCGTAGCT A G C T A C G A C G T C T A

3. Start End

4. 20 30 10 A nomeação é praticamente aleatória no início e no final, onde a chance de erro é alta (baixo valor de Phred) O programa Phred lê o cromatograma e nomeia as bases cromatograma acgatctcgctagctgctactgtagccgcgattattcgcgatctacgtatatcgcgatcgatc Cada base tem uma chance de erro de sua nomeação (10% = 0,1) A escala de Phred é semelhante à de pH multiplicado por 10: - chance de erro de 0,001 = 10 -3 = Phred 30 Processamento de seqüências

5. In the Pursuit of Optimal Sequence Trimming Parameters for EST Projects Fabiano C. Peixoto & J. Miguel Ortega LCC-CENAPAD A T G C BIOINFORMÁTICA UFMG

6. Noticed: BLAST results Phred 15 Too much trimming 0 10 20 30 40 50

7. Query: 469 TTAGGAGGATCGTTTTTAGAATCCCCTGCAACGTTACCACGGTGGATTTCACTGACTGCG 528 |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||| Sbjct: 1038 ttaggaggatcgtttttagaatcccctgcaacgttaccacggtggatttcactgactgcg 979 Query: 529 ACGTTCTTAACGTTGAATCCAACGTTGCTACCAgggagagcctcagtaagtgcttcatga 588 ||||||||||||||||| || |||||||||||||||||| |||||||||||||||||||| Sbjct: 978 acgttcttaacgttgaagcccacgttgctaccagggagaccctcagtaagtgcttcatga 919 Query: 589 tgcatttcgacagaattgacttcagtcgacaaaccttgcggagcaaaagtgacgaccata 648 |||||||||||||| |||||||||| |||| ||||||||||| ||||||||||||||||| Sbjct: 918 tgcatttcgacagacttgacttcagccgaccaaccttgcggaccaaaagtgacgaccata 859 Query: 649 ccaggcttgatgataccagtttcaacgc 676 |||||||||||||||||||||||||||| Sbjct: 858 ccaggcttgatgataccagtttcaacgc 831.TGAAGCTTTCAGCTTCTTTAGGAGGATCGTTTTTAGAATCCCCTGCAAC GTTACCACGGTGGATTTCACTGACTGCGACGTTCTTAACGTTGAATCCAA CGttGCTACCAgggagagcctcagtaagtgcttcatgatgcatttcgaca gaattgacttcagtcgacaaaccttgcggagcaaaagtgacgaccatacc aggcttgatgataccagtttcaacgcctcggggccaggctggcgtgaaca gggcctagcgggtccgcgggggaagggtcccggctcaatccaccaataga gcggagctaaagtgacgggggcgcca Phred 15

8. Experimental approach Sequences: pUC18 plasmidial vector (published sequence) Sequence reaction: Single pool - 3 plates (96 samples) MegaBACE sequencer 3 reads for each plate, esd processing - 846 reads Processing: BLAST (MegaBLAST, as in UniGene) Phred trim: a chromatogram analyzer trim_alt: trim_cutoff parameter 1% up to 25%

10. 16%17% Trim_alt sequence BLAST gaps/missmatches (% of bases) Additional bases 3%