Reagent for diagnosing chronic myeloid leukemia blast crisis based on hnRNPA1 splicing variant and kit
A technology of variants and reagents, applied in the field of reagents and kits for diagnosing CML blastic phase based on hnRNPA1 splicing variants, can solve problems such as lack of alternative splicing, and achieve high clinical application value, high sensitivity and specificity Effect
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[0032] 1. Sample collection of CML patients and healthy controls
[0033] According to the NCCN diagnostic criteria for CML in 2016, peripheral blood samples were collected from patients with chronic myeloid primary and blast disease who were not treated for the first time, as well as peripheral blood samples from the healthy physical examination group.
[0034] 2. Extraction of peripheral blood mononuclear cells
[0035] Ficoll density gradient centrifugation method: ①Put 5ml whole blood into a 50ml centrifuge tube, dilute with 5ml PBS solution, and mix gently; ②Take two 15ml centrifuge tubes and add 5ml Ficoll solution first. Then gently add the diluted blood to the upper layer of Ficoll with a pipette (5ml each) (to avoid mixing the two solutions); ③centrifuge at 2000rpm for 20min; ④use a pipette to draw the white cell layer (ie PBMC) into a clean 15ml ⑤Add PBS to 10-15ml, centrifuge at 1500rpm for 10min, remove the supernatant, add Trizol and mix well. Store at -80°C for...
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