Normalized RNA-Seq of Ipomoea batatas roots cultured in absence of nitrogen
One total RNA sample from Ipomea batatas. The cDNA synthesis from the poly(A)+ RNA that were isolated from total RNAs. Firststrand cDNA synthesis was primed with a N6 randomized primer. 454 adapters A and B were ligated to the 5' and 3' ends of the cDNA. The cDNA was finally amplified with PCR. Normalization was carried out by one cycle of denaturation and reassociation of the cDNA, resulting in N1-cDNA. Reassociated ds-cDNA was separated from the remaining ss-cDNA (normalized cDNA) by passing the mixture over a hydroxylapatite column. After hydroxylapatite chromatography, the ss-cDNA was PCR amplified.