| Library |
| Library name |
Construction protocol |
Strategy |
Source |
Selection |
Layout |
|
After the RNA sample has passed quality control, mRNA from eukaryotic organisms is enriched using magnetic beads with Oligo(dT). Fragmentation buffer is then added to break the mRNA into short fragments. Using these mRNA fragments as templates, first-strand cDNA is synthesized using random hexamers. Subsequently, second-strand cDNA is synthesized by adding buffer, dNTPs, DNA polymerase I, and RNase H. The double-stranded cDNA is purified using AMPure XP beads. The purified double-stranded cDNA undergoes end repair, adenylation, and adapter ligation, followed by size selection using AMPure XP beads. PCR amplification is then performed, and the PCR products are purified using AMPure XP beads to obtain the final library. After library construction, initial quantification is conducted using Qubit 2.0. The library is diluted, and the insert size of the library is assessed using the Agilent 2100. Once the insert size meets the expected range, the effective concentration of the library is accurately quantified using a Q-PCR method to ensure library quality. |
RNA-Seq |
TRANSCRIPTOMIC |
RANDOM PCR |
PAIRED
|
|
