GSM9793169: Riboseq_High_1; Mus musculus; OTHER
Sample was treated with specific lysis buffer with cycloheximide (50mg/mL) to acquire the lysate. Following PAGE purification, the both ends of RPF were phosphorylated and ligated with 5' and 3' adapters respectively. The RNA samples were then treated with rRNA depletion kit（Qiagen,334387） to deplete the samples of as much rRNA contamination as possible. Then the fragments were reversely transcribed to the cDNAs and amplified by PCR[1-2].After library construction（Multiplex Small RNA Library Prep Set for ILLumina (Set1)，NEB，E7300L）,the concentration of library was measured by The Qubit® 2.0 Fluorometer and adjusted to 1ng/μL. Agilent 2100 Bioanalyzer was deployed to examine the insert size of the acquired library. At last, the accurate concentration of cDNA library was again examined using qPCR. Once the insert size and concentration of the library was correct, the samples can then be subjected for sequencing.