454 sequencing of cDNAs derived from total seedling mRNA
A cDNA library was constructed from 10 seedlings from 10 elite C. japonica trees in Japan. After total RNA was extracted from each seedling using a CTAB-based method, each batch of RNA was mixed equal-molarly. cDNA synthesis was carried out using SMART cDNA construction kit (Clontech) and normalized using a cDNA Normalization Kit (Evrogen) by Dragon Genomics Centre, TAKARA BIO Inc. (Yokkaichi, Japan). The library was pyrosequenced by Roch 454 GS FLX Titanium in one and a quarter pico-tirer plate.
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