Computational protocol: Construction and Evaluation of Normalized cDNA Libraries Enriched with Full-Length Sequences for Rapid Discovery of New Genes from Sisal (Agave sisalana Perr.) Different Developmental Stages

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[…] Vector-derived and ambiguous sequences were eliminated using online software (vecscreen) from NCBI. The EST sequences were clustered and assembled into contiguous consensus sequences (contigs) using the program Seqman and Megalign of DNAstar software. The non-redundant sequences were searched against protein databases obtained from NCBI with a search threshold of E < 1.0 × 10−5. All similarity searches were executed using the BlastN, BlastX, and tBlastX tools. Phylogenetic trees were constructed by the Neighbor-Joining (NJ) method using the NJ algorithm implemented in the Molecular Evolutionary Genetics Analysis (MEGA) software version 5.0. The Blast results were used to obtain further information on the function and motif through the InterPro member databases []. […]

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