Degradome sequencing of of Nelumbo nucifera druing seed germination
Approximately 150 ng of poly(A)+ RNA was used as input to the SOLiD whole transcriptome analysis kit, following the manufacturer’s instructions except that (1) the initial RNaseIII-catalyzed RNA fragmentation was omitted and (2) a large size range was gel isolated after the RT-PCR. Omitting the RNaseIII fragmentation step restricts the initial adapter ligation to only those RNAs containing 5'-monophosphates. Libraries were equenced using the P1 (5') adapter only, resulting in the sequencing of the first 35 nucleotides of the inserts that represented the 59 ends of the original RNAs.