Snippet: , followed by neutralization of the sample with 500 mM Tris (pH 7.4). SUBCELLULAR FRACTIONATION 293T cells grown in 100 mm dishes were transfected with 30 μg of plasmid DNA, 60 μl of TurboFectin (Origene, #TF81005) and 750 µL of Opti-MEM. Cells were trypsinized after 24 h, washed with PBS, and collected by centrifugation at 2000 × _g_ for 5 min. The pellets were resuspended in 400 μl of Buf

Snippet: Others have reported successful transfection of HAP1s with TurboFectin 8.0 (OriGene #TF81005), although we typically observe significantly reduced efficiency compared;classified as pathogenic or likely pathogenic in ClinVar Table 2: Plasmid DNA amounts for co-transfection in epegRNA architecture screen. We typically use the TurboFectin 8.0 transfection reagent (OriGene #TF81005), but an alternative such as Lipofectamine3000 is suitable if it efficiently transfects HEK 293T cells. …

Snippet: co-transfection of the epegRNA-containing pU6-tevopreq1-GG-acceptor, pCMV-PEmax-P2A-GFP, and pEF1a-hMLH1dn plasmids into HEK293T cells was carried out using the TurboFectin 8.0 transfection reagent (OriGene #TF81005). 100,000 cells were seeded 24 hours before transfection in 24-well plates. Plasmids were co-transfected in the following amounts: 75 ng (pU6-tevopreq1-GG-acceptor), 263 ng (pCMV-PEma

Snippet: Fetal Bovine Serum BioWest S1400 TurboFectin 8.0 Origene TF81005 Lipofectamine3000 Thermo Fisher Scientific 15292465

Snippet: Then, the cells were transduced using OriGene TurboFectin 8.0 Transfection Reagent (Cat # TF81005)