Ccl 243

Manufactured by Thermo Fisher Scientific

The CCL-243 is a laboratory centrifuge designed for general-purpose applications. It features a compact and efficient design, providing reliable performance for routine sample processing tasks.

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3 protocols using ccl 243

Single-Cell DNA Extraction and Staining

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K562 cells (ATCC^® CCL-243™) were cultured in high glucose Dulbecco's modified Eagle's medium (DMEM) (Gibco) supplemented with 10% Fetal Bovine Serum (FBS) (Gibco) and 1% penicillin-streptomycin (Gibco). HepG2 cells (ATCC^® HB-8065™) were cultured in low glucose (1 g/l) DMEM (Gibco), 1% Glutamax (Gibco), 1% non-essential amino acids (NEAA) (Gibco), 10% FBS (Gibco) and 1% penicillin–streptomycin (Gibco). H1 ES cells was a gift from John Chua's lab. The cells were harvested, and the DNA was extracted using DNeasy blood and tissue kit (QIAGEN) for the low-input assays. DNA concentrations were quantified using Qubit dsDNA HS Assay Kit (Thermo Scientific) in Qubit 3.0 (Invitrogen). The cells were stained with CellTrace Calcein Red-Orange AM (Life Technologies), diluted in Phosphate Buffered Saline (PBS) (Gibco) to 1 cell/μl concentration and isolated in 0.2 ml PCR tubes for the single-cell assay. The tubes were observed under the microscope to confirm the presence of a single fluorescently stained cell.

Viswanathan R., Cheruba E, & Cheow L.F. (2019). DNA Analysis by Restriction Enzyme (DARE) enables concurrent genomic and epigenomic characterization of single cells. Nucleic Acids Research, 47(19), e122.

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Polystyrene Microspheres and K562 Cell Culture

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10.2 μm yellow-green polystyrene (PS) microspheres (Magsphere Inc.) were mixed in deionized (DI) water to an approximate concentration of 1 × 10⁶ mL⁻¹. K562 cells (American Type Culture Collection (ATCC®) CCL-243^™) were cultured in RPMI 1640 (Gibco, Life Technologies) medium containing 1% penicillin/streptomycin (PIS) and 10% fetal bovine serum (FBS). The K562 cells were maintained in 5% CO₂, 37 °C cell culture incubators (HERAcell VIOS 160i CO₂ Incubator, Thermo Fisher Scientific). Since K562 cells were non-adherent cells and would not clump, there was no need to use Trypsin to resuspend cells. Before each experiment, the RPMI 1640 medium was replaced, and cells were diluted to an approximate concentration of 1 × 10⁷ mL⁻¹. The suspension was injected into the chamber using a 1 mL syringe (309659, Becton Dickinson).

Kang P., Tian Z., Yang S., Yu W., Zhu H., Bachman H., Zhao S., Zhang P., Wang Z., Zhong R, & Huang T.J. (2020). Acoustic tweezers based on circular, slanted-finger interdigital transducers for dynamic manipulation of micro-objects. Lab on a chip, 20(5), 987-994.

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Culturing Human Cell Lines

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Cells were maintained in medium supplemented with 1X antibiotic-antimycotic (Gibco, 15240062), and 10% fetal bovine serum (Corning, 35-011-CV) in 5% CO2 at 37 °C. Human foreskin fibroblasts immortalized with hTERT (BJ-5ta, ATCC® CRL-4001™) were cultured in Dulbecco’s Modified Eagle’s Medium (Sigma-Aldrich, D5796). Human Lymphoblast K-562 (ATCC® CCL-243™) were cultured in RPMI1640 medium (Gibco, 11875093) additionally supplemented with 3 mg/L puromycin (Sigma, P8833). Expi293F cells (Gibco, A14527) were cultured in Expi293 medium (Gibco, A1435102).

Jo M.H., Li J., Jaumouillé V., Hao Y., Coppola J., Yan J., Waterman C.M., Springer T.A, & Ha T. (2022). Single-molecule characterization of subtype-specific β1 integrin mechanics. Nature Communications, 13, 7471.

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