GSM6246342: CP_273020; Homo sapiens; RNA-Seq
After the samples are qualified, the total RNA library preparation and library inspection are carried out according to the following experimental steps: (1) rRNA removal: Ribo-zero kit was used to remove rRNA, followed by high temperature and metal ions. (2) Synthesis and purification of cDNA: using RNA as a template, a six-base random primer is used to synthesize a strand of cDNA,Then add buffer, dNTPs, and enzyme to synthesize double-stranded cDNA, and then use VAHTSTM DNA Clean Beads Purification of double-stranded cDNA; (3) Fragment screening: the purified double-stranded cDNA is first subjected to end repair, A-tail, and sequencing ligation  head, followed by fragment screening with VAHTSTM DNA Clean Beads; (4) PCR enrichment: UDG enzyme degrades the second strand of cDNA containing U, and performs PCR enrichment, Finally, use VAHTSTM DNA Clean eads to purify the PCR product; (5) Library inspection: use Qubit to accurately quantify the library concentration, and Agilent 2100 Bioanalyzer to determine Library fragment size distribution to assess suitability for onboarding. Ribo-zero kit for rRNA removal;One-stranded cDNA was synthesized using RNA as a template and six-base random primers; The second-stranded cDNA was synthesised by adding buffer, dNTPs and enzymes, followed by purification of the double-stranded cDNA using VAHTSTM DNA Clean Beads. purification of double-stranded cDNA;Accurate quantification of library concentrations with Qubit, determination of library fragment size distribution with Agilent 2100 Bioanalyzer