Name: GSM7165705
Instrument: Illumina NovaSeq 6000
Strategy: ncRNA-Seq
Source: TRANSCRIPTOMIC
Selection: size fractionation
Layout: PAIRED
Construction protocol: Fresh tumor samples and the paired adjacent normal tissue were collected and treated with specific lysis buffer with cycloheximide. Concentration of the lysate was measured by the NanoDrop™ 2000 Spectrophotometer. Then the lysate was treated with unspecific endoribonuclease RNase I to digest RNA other than ribosome protected fragments (RPFs) and monosomes were isolated by size-exclusion chromatography with MicroSpin S-400 HR columns. The RNA samples were then treated with rRNA depletion kit to eliminate as much rRNA contamination as possible. Subsequently, the relatively short (20~38 nt) RPFs were purified using PAGE and the both ends of RPF were phosphorylated and ligated with 5' and 3' adapters respectively. Then the fragments were reversely transcribed to the cDNAs and amplified by PCR.