To perform the cytogenetic analysis, the cells were cultured in T75 cm 2 flasks until reaching 70-80% confluence, thus ensuring the maintenance of the exponential phase of growth without reaching the stationary phase. Metaphases were obtained by adding colcemid (KaryoMAX ™ Colcemid ™ solution in HBSS, Gibco, 15,210-040, USA) at a final concentration of 0.1 μg/ml, 3 h before the trypsinization. The cultures were then processed according to the conventional air-drying protocol, as previously described (Sirchia and Luparello 2007; (link)Mannino et al. 2019; (link)Caradonna et al. 2020) (link).
Karyomax colcemid solution in
Manufactured by Thermo Fisher Scientific
Sourced in United States
KaryoMAX™ Colcemid™ solution is a cell culture reagent used for metaphase arrest. It contains colcemid, a microtubule-inhibiting agent that blocks cell division at metaphase, enabling the collection of chromosomes for karyotyping and cytogenetic analysis.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using karyomax colcemid solution in
1
Cytogenetic Analysis of Cell Cultures
2
Cytogenetic Analysis of Fibroblast Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After the cells reached 70% confluence, we added
KaryoMAX® Colcemid™ Solution in PBS-10 µg/ml (Gibco,
USA) to each flask to a final dilution of 25 µl/ml, which was
then incubated at 37°C for 45 minutes. We monitored the
changes in cell morphology with an inverted microscope
until the fibroblasts detached. For hypotonic treatment, we
slowly and carefully added 13 ml of 0.056% KCl (Merck,
Germany) with distilled water, followed by incubation at
37°C for 11 minutes, and fixed with methanol: acetic acid
(3:1) solution. In order to obtain G-bands, we aged the slides
at 60°C overnight. We carried out the staining procedure using
Giemsa solution 1:10 (Gibco, USA). Whenever possible, we
analysed 15 metaphases. Before printing out each karyotype
and counting each chromosome by writing a number on each
sister chromatid pair, we observed the slides under a light
microscope at ×10 and ×100 magnifications.
KaryoMAX® Colcemid™ Solution in PBS-10 µg/ml (Gibco,
USA) to each flask to a final dilution of 25 µl/ml, which was
then incubated at 37°C for 45 minutes. We monitored the
changes in cell morphology with an inverted microscope
until the fibroblasts detached. For hypotonic treatment, we
slowly and carefully added 13 ml of 0.056% KCl (Merck,
Germany) with distilled water, followed by incubation at
37°C for 11 minutes, and fixed with methanol: acetic acid
(3:1) solution. In order to obtain G-bands, we aged the slides
at 60°C overnight. We carried out the staining procedure using
Giemsa solution 1:10 (Gibco, USA). Whenever possible, we
analysed 15 metaphases. Before printing out each karyotype
and counting each chromosome by writing a number on each
sister chromatid pair, we observed the slides under a light
microscope at ×10 and ×100 magnifications.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!