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E randomly selected and the sizes of the inserts in these clones were assessed after digestion with appropriate restriction enzymes. These insert sizes ranged from 400 to 1,600 base pairs (bp) with an average of 1.1 kilobase pairs (kb). A total of 5,472 cDNA clones were subsequently randomly isolated and sequenced from the 5' end in order to generate ESTs. The sequences were trimmed of vector sequence, adaptor sequence, poly(A) tail and low-quality sequence and filtered for minimum length (150 bp), resulting in a total of 5,076 high quality ESTs. The average length of submitted ESTs was 468 nucleotides (nt). Cluster formation and analysis To identify overlapping EST sequences, improve base accuracy and transcript length, and to produce nonredundant EST data for further functional annotation and comparative analysis, the 5,076 ESTs from the A. avenae library were grouped by sequence identity into clusters. Based upon regions of nucleotide identity, EST sequences were merged into contiguous consensus sequences (contigs). 'Contig' member ESTs derive from identical transcripts while 'cluster' members may derive from the same gene but represent different transcript splice isoforms (i.e ESTs form contigs, contigs form clusters). Two thousand seven hundred non-redundant EST clusters were generated from the ESTs (Table 1). In 2,005 cases, clusters consist of a single EST, whereas the largest single cluster contains 81 ESTs (1 case) (Fig. 2). The majority of contigsTable 1: A. avenae cDNA library, ESTs and clusters statisticsTitre of cDNA library (pfu/ml) Average cDNA insert size Total cDNA clones picked and sequenced Sequences passing quality check Average length Singletons Contigs Total number of clusters1.2 ?105 1.1 kb 5472 5076 (93 ) 468 bp 2005 695Page 3 of(page number not for citation purposes)BMC Genomics 2009, 10:http://www.biomedcentral.com/1471-2164/10/Histogram showing the distribution of ESTs from A. avenae by cluster size Figure 2 Histogram showing the distribution of ESTs from A. avenae by cluster size. For example, there were five clusters of size 23 containing a sum total of 115 ESTs. Distribution of contig sizes is not shown.Table 2: The most abundantly represented transcripts in the A avenae cDNA libraryNon-redundant GenBank No. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24a C.Cluster ID AAC00129 AAC00161 AAC00061 AAC00128 AAC00023 AAC00091 AAC00187 AAC00105 AAC00123 AAC00255 AAC00048 AAC00088 AAC00109 AAC00321 AAC00541 AAC00269 AAC00121 AAC00143 AAC00163 AAC00313 AAC00112 AAC00295 AAC00024 AAC00265 AACESTs 81 73 71 64 57 57 32 27 24 24 23 23 23 23 23 19 18 18 17 17 PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/16989806 16 16 15 15Best identity descriptor Actin [Panagrellus redivivus] Hypothetical protein F42A10.7 [C. elegans] Small heat shock protein OV25-2 [Onchocerca volvulus] Collagen family member (col-144) [C. elegans] Troponin C [C. Lenalidomide brenneri] Tropomyosin [Heligmosomoides polygyrus] Cuticular collagen [Teladorsagia circumcincta] Hypothetical protein CBG24046 [C. briggsae] Novel Novel Temporarily assigned gene name family member [C. elegans] Novel Hypothetical protein CBG15446 [C. briggsae] Novel SXP/RAL-2 family protein [Anisakis simplex] Hypothetical protein CBG02561 [C. briggsae] Cytochrome c oxidase subunit I [Steinernema carpocapsae] Cuticle preprocollagen [Meloidogyne incognita] Phosphoenolpyruvate carboxykinase [Haemonchus contortus] ATP synthase subunit family member (atp-2) [C. elegans] Cytochrome c oxidase subunit III [Ascaris suum] Novel Hypothetica.