Fig. 1From: An improved method of constructing degradome library suitable for sequencing using Illumina platformThe scheme for constructing improved degradome library. For sequencing purposes, the degradome library generated by this method can be treated as small RNA library and the resulting reads are ~ 27 nt long. The procedure includes: (1) poly(A) RNA isolation; (2) 5′RNA adapter ligation to uncapped poly(A) RNA with 5′ monophosphate; (3) reverse transcription to generate 1st strand cDNA using an oligo(dT)-tailed adapter (RT-primer); (4) second strand synthesis (1st PCR amplification); (5) EcoP15I digestion to generate ~ 27 nt long reads; (6) ligation of EcoP15I digestion products with a 3′ds-DNA adapter; (7) purification of ligation products on a PAGE gel; (8) degradome library enrichment (2nd PCR amplification); (9) purification of the final product on a PAGE gel; (10) library pooling and sequencing using Illumina HiSeq platformBack to article page