Small RNA Solexa sequencing of multiple tissues of papaya, Carica papaya. Carica papaya

The tissues of male flowers, female flowers and hermaphrodite are collected and stored in liquid nitrigen immediately. Total RNA was isolated from male, hermaphrodite, and female flowers using the Trizol reagent (Invitrogen USA). Low molecular weight RNA was enriched from the total RNA by precipitating with 0.4M NaCl and polyethylene glycol (PEG). The enriched low molecular weight RNA was then separated on a denaturing 15% polyacrylamide gel. The band corresponding to the RNA standard of 18-30 nucleotides was then excised from the gel and eluted overnight in 0.4M NaCl at 4 degrees Celsius, and subsequently ligated with 5’ and 3’ Illumina small RNA adapters. The adapter ligated library was converted into cDNA and amplified by using PCR and sequenced by Illumina Genome Analyzer II. Overall design: Examination of small RNA transcriptomes in multiple tissues of papaya using Illumina Genome Analyzer II sequencer.