Preparation method of sample used for electrophoresis imaging analysis of single-cell gelatin and reagent kit
A single-cell gel and image analysis technology, which is applied to the sample preparation of single-cell gel electrophoresis image analysis, and the field of kits for implementing this method, can solve problems affecting popularization and application, threats to user health, difficult disposal of experimental waste, etc. problem, to achieve the effect of improving sensitivity and image quality, safe and reliable use, and high success rate
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Embodiment 1
[0025] (1) Preparation of lymphocyte samples. 2ml of venous blood was drawn from young healthy male volunteers with no smoking history and no recent medication history, and anticoagulated with heparin. Then draw the diluted blood and add it to the lymphocyte separation medium (1:1), and centrifuge at 2000r / m for 20min. Aspirate the lymphocyte layer, wash twice with PBS, 1500r / m, 10min. The precipitate was diluted into a suspension with RPMI-1640 and counted (1×107 / mL). Then with H 2 o 2 Exposure: Divided into 6 dosage groups, H 2 o 2 The final concentrations were 50 μM, 100 μM, 150 μM, 200 μM, 400 μM, and 800 μM, respectively. A blank control group (group 0) was set up and placed in a 37°C water bath for 1 hour to prepare lymphocyte samples.
[0026](2) Laying glue. Preheat a frosted glass slide at 40°C, and then evenly coat 1% eutectic point agarose gel prepared with calcium- and magnesium-free phosphate buffer solution (PBS) on the frosted glass slide, and store at lo...
Embodiment 2
[0035] (1) Prepare bone marrow cell samples. Bone marrow cells were obtained from femurs of Kunming mice (Guangzhou Experimental Animal Center) aged 5 to 6 weeks and weighing 22 to 25 g, washed twice with PBS, and then centrifuged at 1500 r / m for 10 minutes. The precipitate after centrifugation was diluted into a suspension with RPMI-1640 and counted (1×107 / mL); 2 o 2 Infected and divided into 6 dose groups, H 2 o 2 The final concentrations were 50 μM, 100 μM, 150 μM, 200 μM, 400 μM, and 800 μM, respectively. A blank control group (group 0) was set up and placed in a water bath at 37° C. for 1 hour to prepare bone marrow cell samples.
[0036] (2) Laying glue. Preheat a frosted glass slide at 41°C, and then evenly coat 1% eutectic point agarose gel prepared with calcium- and magnesium-free phosphate buffer solution (PBS) on the frosted glass slide, and store at low temperature. Cured under the hood to obtain a glass slide covered with the first layer of gel. Preheat 20 μ...
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