GSM8635842: Parp7 siRNA2 2 day Replicate 4; Mus musculus; RNA-Seq
3T3-L1 cells were grown and differentiated, in a 6-well plate, as mentioned above and were transfected with siRNAs targeting Parp7 using the Lipofectamine 3000 Reagent (Invitrogen, L3000015). The cells were collected, and total RNA was isolated using the Direct-zol RNA MiniPrep Plus (ZYMO Research) according to the manufacturer's instructions. The RNA obtained above was used to generate strand-specific RNA-seq libraries. Briefly, the libraries were prepared using the the NEBNext Ultra II Directional RNA Library Prep Kit (New England Biolabs #E7760S) with the NEBNext Poly(A) mRNA magnetic Isolation Module (New England Biolabs #E7490S) for mouse SVF samples and 3T3-L1 cells. Specifically, 0.6µM adapter was used for its ligation and final mRNA library was amplified with 14 x PCR cycles and sequenced on an illumina NEXTseq2000 system by paired-end 50bp using NEXTseq 2000 P3-100cycle sequencing kit (20040559). n= 4 biological replicates.