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Top agar

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0.4% top agar is a semi-solid culture medium used for the cultivation and isolation of microorganisms. It provides a low-concentration gel-like matrix that supports the growth and development of microbial colonies.

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Lab products found in correlation

Difco agar, by BD (1 mentions)

2 protocols using top agar

1

Soft Agar Colony Formation Assay

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Pre-warmed 0.8% bottom agar (Difco™ Agar) (BD, Franklin Lakes, NJ, USA) was prepared with PBS. 1 mL of bottom agar was poured into each well in 12-well plate. Pre-warmed 0.4% top agar (BD) was prepared with culture medium for each cell type. After bottom agar was congealed, 1mL of 0.4% top agar (BD) containing 500 cells of U87MG or 1,000 cells of ahMNCs or hTERT-ahMNCs was poured on the bottom agar. After top agar is congealed, wells were incubated at 37°C and 5% CO2 for 2 weeks. The medium was changed every 3 days.
Lee K.H., Nam H., Jeong D.E., Kim S.S., Song H.J., Pyeon H.J., Kang K., Hong S.C., Nam D.H, & Joo K.M. (2016). Sensitive Tumorigenic Potential Evaluation of Adult Human Multipotent Neural Cells Immortalized by hTERT Gene Transduction. PLoS ONE, 11(7), e0158639.
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2

Soft Agar Assay for Anchorage-Independent Growth

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Soft agar assay was performed as previously described [75 (link)]. Briefly, cells (2 × 104) were suspended in 0.4% top agar (BD Biosciences) and then plated onto 0.8% base agar in six well-plates. The plates were incubated at 37°C with 5% CO2 for 2 weeks. Random fields were chosen and photographed. Colony areas were counted by NIH Image J software. The colony sizes of 20 μm or larger were scored for calculation of the percentage of soft agar colony.
Li W., Wang F., Shi J., Feng Q., Chen Y., Qi X., Wang C., Lu H., Lu Z., Jia X., Yan Q., Gao S.J, & Lu C. (2020). Sperm associated antigen 9 promotes oncogenic KSHV-encoded interferon regulatory factor-induced cellular transformation and angiogenesis by activating the JNK/VEGFA pathway. PLoS Pathogens, 16(8), e1008730.
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