PMC:3475480 / 4946-5826 JSONTXT

{"target":"https://pubannotation.org/docs/sourcedb/PMC/sourceid/3475480","sourcedb":"PMC","sourceid":"3475480","source_url":"https://www.ncbi.nlm.nih.gov/pmc/3475480","text":"Synthesis of siRNA\nCasz1 siRNA oligonucleotides were synthesized by Genolution (Seoul, Korea). The target sequence for Casz1 was 5'-GATGTGATCCGACATTACA-3' (siRNA#1), 5'-CTAGAACTGCTCTGTATGT-3' (siRNA#2), and 5'-CAATTCGGCAAGTAATGGA-3' (siRNA #3), and nonspecific scrambled siRNA was used as a negative control.\nTo examine the efficacy of siRNAs, siRNAs were transfected into B16F10 cells using G-Fectin (Genolution); 1-2 µL of G-Fectin and 20 nM siRNA were added to 50 µL phosphate buffered saline, and the reaction was incubated for 10 min at room temperature (RT). B16F10 cells were used to seed 24-well plates at 5 × 104 cells/well and 30% confluence, and the transfection mixture was added to the plate. The cells were harvested for RNA extraction after 24 h of incubation. Total RNA was prepared using TRIzol (Invitrogen, Carlsbad, CA, USA) per the manufacturer's instructions.","divisions":[{"label":"Title","span":{"begin":0,"end":18}}],"tracks":[]}