Comb transcriptome
Total RNA is extracted from six Pleurobrachia bachei. Library construction starts with total RNA reverse transcribed to cDNA with an oligo dT primer, with second strand then generated. The cDNA is digested with a restriction enzyme then adaptors ligated onto the double-stranded (ds) DNA.  The adaptor ligated cDNA fragments are the amplified by PCR. This product is purified and processed for emPCR and 454 sequencing according to manufactures procedures. Adaptors for library construction: 5' targeted library A 5' GCCTCCCTCGCGCCATCAG 3' B 5' GCCTTGCCAGCCCGCTCAG 3' 3' targeted library A   5'-CCATCTCATCCCTGCGTGTCCCATCTGTTCCCTCCCTGTCTCAG-3' B  5'-CCTATCCCCTGTGTGCCTTGCCTATCCCCGCAGTCGGTACTTTT-3'