F.vescaH4x4_PoolTranscrSMART_NONnorm
The full-length (FL) enriched cDNA library was prepared using SMART method and MINT cDNA synthesis kit (Evrogene) as suggested by the manufacturer. The FL enriched cDNA library for the Illumina-based high throughput sequencing was prepared as described (Filichkin et al., 2009, Fox et al., 2009). Briefly, 5 µg of double-stranded cDNA in a 50 µL of TE buffer (10 mM Tris, 1 mM EDTA; pH  8.0)  was mixed with 750 µL of nebulization buffer [25% (v/v) glycerol in TE buffer] and sheared for 7 min. in a nebulizer (Invitrogen) using compressed nitrogen at 36 psi. Before ligation with Illumina adapters the fragmented cDNA was purified using “QIAquick PCR Purification Kit” (Qiagen) according to the manufacturer protocol.  The cDNA library was  amplified for 9, 11, and 13 cycles. "