454 ESTs

 

            Next generation sequencing (NGS) technology (Varshney et al. 2009), was employed to compile transcriptomic resources ESTs. cDNA prepared at ICRISAT from 15 tissues representing different developmental stages of the Pusa Ageti genotype were pooled, and normalized to minimize redundancy and enhance efficiency of capture of rare transcripts. ICRISAT in collaboration with JCVI sequenced the normalized cDNA pool using 454/FLX sequencing, a next generation sequencing technology that offers higher throughput relative to Sanger sequencing. Of 496,705 sequence reads 287,766(>50%) were longer than 200 bp. Cluster analysis of these sequences, done at NCGR in collaboration with ICRISAT, provided 127, 754 Tentative Unique Sequences(TUSs).

Download 454/FLX ESTs Dataset

     Full Set - 496,705 entries

     Non-redundant Set - 127,754 entries

     Advanced Transcriptome Assembly - 21,434 entries

(based on CAP3 assembly of full set)


            Similarly, NRCPB has employed 454/FLX sequencing on the cDNA pools from two cultivars, Asha and UPAS 120. As a result of this a total of 1,696,724 sequence reads (566 Mb) with an average read length of 333 bp were generated from these two genotypes at NRCPB. Sequence analysis of these genotypes provided 42,000 unique sequences of which 25,000 were common between these two genotypes.

Download 454/FLX ESTs Dataset from Asha, Upas120.

            Together these transcript sequences represent a significant fraction of the pigeonpea transcriptome, and should be a useful resource for marker development as well as gene discovery and functional studies.