title: Pooled normalized library, normal and regenerating radial organ complex of Holothuria glaberrima, 454 pyrosequencing
design: Generation of this library involved cDNA normalization with a TRIMMER cDNA Normalization kit (Evrogen Cat# NK001). First strand cDNA synthesis was performed with SMARTScribe Reverse Transcriptase (Clontech),  the modified primer for first strand cDNA synthesis (5' – AAGCAGTGGTATCAACGCAGAGTACTTTTGTTTCTTTTGTTTCTTGTTTCTTTTVN - 3'), and the SMART IIA Oligonucleotide (Clontech) (5' – AAGCAGTGGTATCAACGCAGAGTACGCGGG – 3). Pre- and post-normalization amplification of cDNA was performed with Advantage 2 Polymerase Mix (Clontech) and Evrogen PCR Primer M1 (5'-AAGCAGTGGTATCAACGCAGAGT-3'). This library was barcoded with MID RL7.