title: RNA-seq of AGS cells infected with Helicobacter pylori
design: 1) totalRNA was processed by mRNA enrichment or rRNA division. MRNA enrichment: magnetic beads with OligodT were used to enrich the mRNA with polyA tail;  RRNA removal: Hybridization with DNA probes   RRNA, RNaseH selectively digests the DNA/RNA hybrid chain, then digests the DNA probe with DNaseI, and the required RNA is obtained after purification.   2) The RNA obtained was segmented by interrupting buffer, and the random N6 primers were reversely transcribed, and then the cDNA two-strand was synthesized to form double-stranded DNA.   3) The synthetic double-stranded DNA end is flattened and phosphorylated at the 5 'end to form A sticky end protruding an "A" at the 3' end, and then connected to A bulbous connector with A protruding "T" at the 3 'end.   4) The ligand products were amplified by PCR with specific primers.   5)PCR products were thermally denatured into single strands, and then the single strand DNA was cycled with a bridge primer to obtain a single strand circular DNA library.