Cells were treated with 60 ng/ml KaryoMAX Colcemid Solution (Invitrogen) for 6 h to arrest the cells in metaphase. Next, the cells were harvested by trypsin treatment. After two washes in PBS, cells were gently resuspended in a hypotonic (75 mM) KCl solution and incubated at 37°C for 20 min. Cells were then centrifuged and resuspended in cold methanol:acetic acid 3∶1 (v/v) for fixation. After fixation, cells were centrifuged again and resuspended in a small volume (200 µl/106 cells) of the same cold methanol:acetic acid solution. These cells were spread onto glass slides, allowed to dry, and stained with KaryoMAX Giemsa Stain (Invitrogen). After washing the slides with water, a minimum of 10 chromosome spreads were analyzed.
Karyomax giemsa stain
Manufactured by Thermo Fisher Scientific
Sourced in United States
The KaryoMAX Giemsa Stain is a laboratory reagent used in the field of cytogenetics. It is a staining solution that is specifically designed to visualize and analyze chromosomes during karyotyping procedures. The stain binds to the DNA in chromosomes, enabling the identification and examination of their structure and number.
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Lab products found in correlation
5 protocols using karyomax giemsa stain
1
Metaphase Chromosome Spread Preparation
2
Chromosome Aberration Analysis in MCF10A Cells
3
Cell Migration and Invasion Assay
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Cell motility was measured using cell culture inserts (8.0-μm pore size) for six-well plates (BD Falcon). Cell invasion was quantitated using BioCoat BD Matrigel invasion chambers (8.0-μm pore size). Cells (2 × 105) were grown in the insert. After 24 h, those cells retained inside the insert were removed, and those that migrated to the other side of the insert were fixed and stained with Karyomax Giemsa stain (Gibco, Invitrogen).
4
Capturing and Sampling Macaques for Malaria Screening
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Trapped macaques were sedated with 0.3–0.5 mL ketamine (Vetoquinol, UK) via intramuscular injection before blood collection (1–5 mL). The blood samples were transferred into ethylenediaminetetraacetic acid (EDTA) tubes (Becton-Dickinson, Franklin Lakes, NJ, United States). Blood spots on filter paper (Whatman® No. 1) were also collected. Thick and thin blood films were also prepared for malaria parasite examination by microscopy (BFMP). Each macaque was marked and then handed over to the Wildlife Department for release into the deep forest or areas of less conflict with humans. The blood samples in EDTA tubes were kept at 4 °C during transportation to the Parasitology Laboratory, IMR. Thin blood smears were fixed with methanol. Both thick and thin blood smears were then stained with 3–10% KaryoMAX® Giemsa Stain (Gibco®, Life Technologies Carlsbad, CA, United States) for 30–60 min, examined and results were recorded. Blood samples in EDTA tubes were subjected to DNA extraction.
5
Retrotransposition Assay in HeLa Cells
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We used HeLa cells (HeLa JM, kindly provided by Dr John Moran, University of Michigan, Ann Arbor) and a retrotransposition reporter, pRTC2, an L1.3-containing version of the retrotransposition reporter plasmid (20 (link)) that we extensively modified as described in detail in the Supporting Information that accompanies Cook et al. (38 (link)) except that the version used here lacks the puromycin N-acetyl-transferase gene (Figure 1C ). HeLa cells were plated in a 6-well dish at 2 × 105 cells/well and within 24 h a mixture of 3 μl FuGene®6 Transfection Reagent (Roche) and 1 μg of p111rtc, p151rtc or p555rtc were applied to the cells (per the supplier's suggestions). After 3 days, 400 μg/ml G418 antibiotic (Gibco) was added to select for G418 resistant foci and incubation was continued for an additional 10–14 days with media change as needed. The cells were then washed twice with 1× PBS, fixed (2% formaldehyde, 0.2% glutaraldehyde in 1× PBS) and stained with Karyo Max® Giemsa Stain (Gibco). After 30 min the stain was removed and the cells were washed repeatedly with 1× PBS until the background was colorless.
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