Small RNA sequences of Meloidogyne incognita: J4 stage
Total RNA was isolated at J4 stage of the nemtode life cycle and from the total RNA, nucleotides of length 18- 30 bp were obtained by gel separation and the purified short nucleotide RNA sequences were ligated with adapters (3' adapter 6-20nt and 5' adapter 12-25 nt) at both ends which was followed by purification  (T4 RNA Ligase, 200 U, 30 U/ul, Takara: D2050). Ligated small RNAs were reverse transcribed with the complimentary sequence of 3' adaptor (SuperScriptH III Reverse Transcriptase, Invitrogen: 18064-014), and amplified. Finally, the PCR amplification products were sequenced at BGI (Beijing Genome Institute) using Illumina HiSeq 2000.