GSM9359964: L125Rho_3w_3; Mus musculus; RNA-Seq
Retinas of 3-week-old C57BL/6J, hRHOL125R/L125R and hRHOP347L/P347L mice (n=3 mice per genotype) were quickly separated and immediately homogenized in TRIzol reagent (Invitrogen 15596018CN). RNAseq library was prepared by Novogene (Beijing, China). Non strand specific libraryMessenger RNA was purified from total RNA using poly-T oligo-attachedmagnetic beads. After fragmentation, the first strand cDNA wassynthesized using random hexamer primers followed by the secondstrand cDNA synthesis. The library was ready after end repair, A-tailing,adapter ligation, size selection, amplification, and purification. Thelibrary was checked with Qubit and real-time PCR for quantification andbioanalyzer for size distribution detection.Strand specific libraryMessenger RNA was purified from total RNA using poly-T oligo-attachedmagnetic beads. After fragmentation, the first strand cDNA wassynthesized using random hexamer primers. Then the second strandcDNA was synthesized using dUTP, instead of dTTP. The directionallibrary was ready after end repair, A-tailing, adapter ligation, sizeselection, USER enzyme digestion, amplification, and purification. Thelibrary was checked with Qubit and real-time PCR for quantification andbioanalyzer for size distribution detection.