Blood samples were taken just before and within a month after each CT examination (Table 1 ). Mononuclear blood cells were isolated from heparinized PB from each sample using BD Vacutainer CPT tubes (BD Biosciences, San Jose, CA, USA) according to the manufacturer’s instructions. Cells were suspended in RPMI 1640 medium (Nacalai Tesque, Kyoto, Japan) containing 20% fetal bovine serum (Equitech Bio, Keilor East, Australia), 2% phytohaemagglutinin-HA15 (Remel, Lenexa, KS, USA) and 60 μg/ml kanamycin solution (Life Technologies, Carlsbad, CA, USA) in a tube. Lymphocytes were cultured in a 5% humidified CO2 incubator at 37°C for 46 h. Then, colcemid solution (Wako, Osaka, Japan) was added (final concentration: 50 ng/ml or 0.05 μg/ml) and cells were cultured for an additional 2 h. After 48 h of culture, chromosome preparations were made according to the standard cytogenetic procedure [11] .
Kanamycin solution
Manufactured by Thermo Fisher Scientific
Sourced in United States
Kanamycin solution is a type of laboratory reagent used in molecular biology and microbiology. It is a concentrated solution of the antibiotic kanamycin, which is commonly used for selection and maintenance of bacterial cultures containing antibiotic resistance genes.
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Lab products found in correlation
5 protocols using kanamycin solution
1
Lymphocyte Isolation and Metaphase Preparation
2
Gamma-Ray Irradiation Protocol for Lymphocyte Analysis
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PB samples were irradiated with gamma-rays (Gamma cell 40, Best Theratronics, Ottawa, Ontario, Canada; installation date: March, 2009.) at eight doses (0, 10, 20, 50, 100, 200, 500 or 1000 mGy). Plastic microtubes containing whole blood were irradiated at a distance of 16 cm at room temperature with gamma-rays from a 60Co radiation source (1.11 TBq) at a dose rate of 26.26t (time: min) + 6.42 mGy per min, where 26.26 is the dose rate and 6.42 is the dose to the sample entering and leaving the irradiation source. The doses were measured using an ionization chamber detector for gamma-rays.
Mononuclear blood cells were isolated from heparinized PB samples using BD Vacutainer CPT tubes (BD Biosciences, San Jose, CA, USA) according to the manufacturer’s instructions. Cells were suspended in RPMI 1640 medium (Nacalai Tesque, Kyoto, Japan) containing 20% fetal bovine serum (Equitech Bio, Keilor East, Australia), 2% phytohaemagglutinin-HA15 (Remel, Lenexa, KS, USA) and 60 μg/ml of kanamycin solution (Life Technologies, Carlsbad, CA, USA) in a 6-well plate. Lymphocytes were cultured in a 5% humidified CO2 incubator at 37°C for 48 h. Colcemid solution (Wako, Osaka, Japan) was added (final concentration: 0.015–0.02 μg/ml) 2 h before cell harvest, then chromosome preparations were made according to a standard cytogenetic procedure [23 ].
Mononuclear blood cells were isolated from heparinized PB samples using BD Vacutainer CPT tubes (BD Biosciences, San Jose, CA, USA) according to the manufacturer’s instructions. Cells were suspended in RPMI 1640 medium (Nacalai Tesque, Kyoto, Japan) containing 20% fetal bovine serum (Equitech Bio, Keilor East, Australia), 2% phytohaemagglutinin-HA15 (Remel, Lenexa, KS, USA) and 60 μg/ml of kanamycin solution (Life Technologies, Carlsbad, CA, USA) in a 6-well plate. Lymphocytes were cultured in a 5% humidified CO2 incubator at 37°C for 48 h. Colcemid solution (Wako, Osaka, Japan) was added (final concentration: 0.015–0.02 μg/ml) 2 h before cell harvest, then chromosome preparations were made according to a standard cytogenetic procedure [23 ].
3
Isolation and Culture of Lymphocytes
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Mononuclear blood cells were isolated from heparinized PBs using BD Vacutainer CPT tubes (BD Biosciences, San Jose, CA, USA) according to the manufacturer’s instructions. Cells were suspended in RPMI 1640 medium (Nacalai Tesque, Kyoto, Japan) containing 20% fetal bovine serum (Equitech Bio, Keilor East, Australia), 2% phytohaemagglutinin-HA15 (Remel, Lenexa, KS, USA), and 60 μg/mL of kanamycin solution (Life Technologies, Carlsbad, CA, USA) in a 15-mL Falcon tube. Lymphocytes were cultured in a 5% humidified CO2 incubator at 37 °C for 48 h. First-division metaphase cells were obtained by treating the culture with colcemid (final concentration, 0.05 μg/mL; Life Technologies) for 48 h. For samples younger than 18 years, colcemid was added for the last 24 h to prevent overcontraction.
4
Lymphocyte Culture and Metaphase Preparation
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Heparinized PB from each patient before and after (within 3–28 days of) the CT scan, and mononuclear blood cells were isolated using BD Vacutainer CPT tubes (BD Biosciences, San Jose, CA, USA) according to the manufacturer's instructions. Cells were suspended in RPMI 1640 medium (Nacalai Tesque, Kyoto, Japan) containing 20% fetal bovine serum (Equitech Bio, Keilor East, Australia), 2% phytohemagglutinin-HA15 (Remel, Lenexa, KS, USA) and 60 μg/ml of kanamycin solution (Life Technologies, Carlsbad, CA, USA) in a 15-ml Falcon tube. Lymphocytes were cultured in a 5% humidified CO2 incubator at 37°C for 48 h. First-division metaphase cells were obtained by treating the culture with colcemid (final concentration, 0.05 μg/ml; Life Technologies) for 48 h.
5
Lymphocyte Cell Culture and Metaphase Preparation
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Heparinized PBs from each patient before and after (within 3–28 days) the CT scan, and mononuclear blood cells were isolated using BD Vacutainer CPT tubes (BD Biosciences, San Jose, CA, USA) according to the manufacturer’s instructions. Cells were suspended in RPMI 1640 medium (Nacalai Tesque, Kyoto, Japan) containing 20% fetal bovine serum (Equitech Bio, Keilor East, Australia), 2% phytohaemagglutinin-HA15 (Remel, Lenexa, KS, USA), and 60 μg/ml of kanamycin solution (Life Technologies, Carlsbad, CA, USA) in a 15-ml Falcon tube. Lymphocytes were cultured in a 5% humidified CO2 incubator at 37 °C for 48 h. First-division metaphases were obtained by treatment with colcemid (final concentration, 0.05 μg/ml; Life Technologies) for 48 h.
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