Gene expression analysis of Q biotype adult whitefly
1. mRNA is captured using the magnetic oligo(dT) beads from the extracted total RNA and first- and second-strand cDNA is synthesized.  2. Bead-bound cDNA is subsequently digested with restriction enzyme NlaIII (recognition sequence CATG).  3. The fragments apart from the 3’cDNA fragments connected to oligo(dT) beads are washed away and a special adapter containing a restriction site for MmeI is ligated to the sticky 5’end of the digested bead-bound cDNA fragments.   4. The MmeI cuts 17 bp downstream from the recognition site, then product the 21 bp tag starting with the recognition site of restriction enzyme NlaIII (CATG).  5. A second adapter is ligated at the site of MmeI cleavage. After enriched by polymerase chain reaction (PCR) amplification, the adapter-ligated tags are purified by 6% tris borate (TBE) buffer, polyacrylamide gel electrophoresis (PAGE).   6. The adapter-ligated tags are fixed on the Solexa Sequencing Chip ( flowecell) to form single-molecule cluster, and then sequencing by synthesis (SBS) is performed to generate millions of raw data according to the manufacturer’s protocols.  7. The raw reads is used for advanced analysis, after dealing with image analysis, base calling and some other filtrations.