Target/Analyte:
Chromosome

Short Name Chromosome
Description Chromosome

Method/Kits with this Target/Analyte
Method/Kit Description
Cytogenetic analysis Giemsa banded (G-banded) karyotyping analysis Cytogenetic analysis Giemsa banded (G-banded) karyotyping analysis was performed for the chromosome analysis of immortalized cells. Cell samples were prepared when cells were in log phase, undergoing active division. The day before the collection, the cells were seeded at ∼40% of confluency in T75 flasks. AEC: 1,400,000/T75; AMC and CMC: 850,000; CTC: 1,200,000; and DEC: 700,000 cells were seeded to T75 flasks overnight. The next day, the cells were treated with 10 μL of KaryoMAX™ Colcemid™ (Gibco™, Cat. No: 15210040, 10 ug/mL)/10 mL complete media for 1 h in a 37°C, 5% CO2 incubator. Then, all cells (including floating cells) were collected and centrifuged at 500 rpm for 7 min at RT. The supernatants were carefully removed, and the pellets were resuspended in 8 mL 0.075 M KCL hypotonic solution for 7 min at RT, then centrifuged again at 500 rpm for 7 min at RT. The supernatants were carefully removed, and the pellets were slowly resuspended in fixation solution (3:1, methanol: acetic acid) for 15 min at RT. The fixative washing step was repeated two times. Then, the cells were stored in a fixative at 4°C until being shipping to Creative Bioarray LLC., Shirley, NY, USA, where G-banded karyotyping services were conducted. In some cases, live cells were shipped at 40% confluency in the Petaka G3™ Cell Culture Cassette (Gemini Bio, West Sacramento, CA, Cat. No: LTPTK25) at RT. Cytogenetic analysis was performed on 20 G-banded metaphase spreads for each cell line.

Studies Utilizing this Target/Analyte
Study Start Date Study Types Description
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