The denaturing solution was replaced with a digestion buffer using a custom-designed 96-well size-exclusion plate for desalting.
Viable cells were established by Trypan Blue exclusion and plated at 5×10 cells/ml in ProN media supplemented with 20 ng/ml EGF to obtain secondary neurospheres.
For trypan blue dye exclusion, 24-well plates were used.
After centrifugation (2,000 × g for 10 min), cell viability was determined by trypan blue exclusion, and cells were plated on 96-well plates, 6-well plates, or 60-mm dishes, depending on the assay being tested.
Afterward, cells were trypsinized, counted by trypan blue exclusion and re-plated (500 cells/well) in duplicate in six-well plates.
Viability was determined by trypan blue exclusion, before cells were plated.
The cells were counted with trypan blue exclusion and 5x10 cells were plated onto low adherent 6-well plates for an additional 2 weeks.
Embryos electroporated with cHairy2 in the ventral midline at HH10 and harvested at HH17 displayed a cell-autonomous exclusion of the mature floor plate marker ARX in targeted cells (n=3 embryos, Fig. 4A-B′) with a concomitant upregulation of Nkx2.2 in some cases, indicating a fate change to P3 identity (Fig. 4A-B′).
For growth curve analysis and Trypan blue exclusion test, ovarian cancer cells were plated in 24-well plates (2 × 10 or 10 × 10 cells cm−2).
A Biomek 2000 workstation (Beckman Coulter, Inc., Fullerton, CA, USA) was used to purify the PCR products by a vacuum-based, size exclusion separation through Montage PCRμ96 Plates (Millipore Corporation, Billerica, MA, USA), following the manufacturer's instructions.
After three washings in ice-cold PBS pH 7.2, PBMCs were counted in a Neubauer plate with trypan blue exclusion of dead cells.
