Genome-wide characterization of long non-coding RNAs and their potential roles in Nicotiana tabacum var. Samsun NN after Tobacco mosaic virus (TMV) infection at different temperatures
Total RNA was extracted using TRIzol regent (Invitrogen, USA) and the first strand of cDNA was synthesized by reverse transcription. The dTTP in the second strand cDNA synthesis reaction buffer was replaced with dUTP, and the second strand of the U-containing cDNA was degraded using the USER enzyme (New England Biolabs, UK). Thus retaining the first cDNA information for PCR extension and obtaining the library.