Comparative Analysis of MicroRNAs between Sporophyte and Gametophyte of Porphyra yezoensis
Figure 1
A flow chart of the procedure for sample preparation and sequencing and for the processing of reads. (a) A flow chart of the procedure for sample preparation and sequencing. (1) Sporophytes and gametophytes of P. yezoensis were frozen rapidly in liquid nitrogen and stored at −80°C before RNA extraction. (2) Total RNA was extracted using the Trizol reagent method. (3) RNA 18–28 nt fragments were gel-purified. (4) A 3′ adaptor was ligated to the 3′ end of sRNAs. (5) A 5′ adaptor was ligated to the 5′ end of sRNAs. (6) sRNAs were amplified by RT PCR. (7) Sequencing. (b) A flow chart of the procedure for processing reads; the numbers in parentheses represent the total reads from PYF and PYL, respectively. (1) Initial processing: remove adapter, filter out low-quality tags, and clean up tags smaller than 18nt. (2) Common/specific tags identified between samples. (3) Length distribution analysis of clean reads. (4) Clean reads matched to P. yezoensis EST sequences using SOAP [25]. (5) Clean reads compared to noncoding RNAs from GenBank and Rfam. (6) siRNA identified. (7) Plant miRNA homologs identified. (8) Annotated sRNAs. (9) miRNA identified by hairpin structure filtering. (10) Target prediction.
