Preparation method of marrow G-band for karyotype analysis
A technology of chromosome karyotype and chromosome, applied in the direction of biochemical equipment and methods, analysis materials, preparation of test samples, etc., can solve the problems of high cost, poor dispersion, non-uniformity, etc., and achieve high accuracy and wide application Foreground, with clear effect
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Embodiment 1
[0039] Example 1 Preparation of bone marrow cell culture termination solution
[0040] The bone marrow cell culture termination solution includes 1.5-5.5 mg / ml ethidium bromide and 8-15 μg / ml colcemid.
[0041] The preparation method of ethidium bromide and colcemid can be prepared by the method described in this example, or by other methods in the art.
[0042] A Preparation of ethidium bromide
[0043] a. Prepare storage solution (concentration is 9mg / ml)
[0044] Add 0.9 g of ethidium bromide to 100 ml of distilled water, stir magnetically for several hours to ensure complete dissolution, then wrap the container with aluminum foil or transfer to a brown bottle, and store at room temperature.
[0045] b. Prepare working solution (concentration is 3mg / ml)
[0046] The stock solution is 1:2 (EB:ddH 2 O) was diluted to a working solution with a concentration of 3mg / ml.
[0047] B formulated colcemid
[0048] a. Prepare storage solution (concentration is 120μg / ml)
[0049...
Embodiment 2
[0052] Example 2 Preparation of Bone Marrow Chromosome Specimens
[0053] In this embodiment, bone marrow cell chromosome samples were prepared as follows. Include the following steps:
[0054] (A) inoculation: the bone marrow cells are inoculated into the bone marrow cell culture medium;
[0055] (B) Termination of culture: adding the termination solution described in Example 1 to the medium of step (A);
[0056] (C) collecting bone marrow cell culture for chromosome preparation;
[0057] (D) Chromosomal specimen preparation: Chromosomal specimens with G-banding were obtained after dyeing.
[0058] In this example, based on the amount of medium used as 5 ml, 50 μl of ethidium bromide with a concentration of 1.5-5.5 mg / ml and 25 μl of colcemid with a concentration of 8-15 μg / ml were added.
[0059] When the culture was terminated in this example, ethidium bromide and colcemid were added to the culture medium, and after shaking evenly, 37°C, 5.0% CO 2 Incubate for 1 hour i...
Embodiment 3
[0085] Embodiment 3 comparative experiment
[0086] The cultured bone marrow cells of the same sample were taken for comparative experiments. Adopt the method of embodiment 1 and embodiment 2 to set up experimental group 1, experimental group 2 and experimental group 3, three groups of experimental groups respectively adopt the cell culture termination solution of different concentrations, wherein:
[0087] The stop solution formula of experimental group 1 is:
[0088] Ethidium Bromide 1.5mg / ml
[0089] Colcemid 8μg / ml
[0090] The stop solution formula of experimental group 2 is:
[0091] Ethidium Bromide 5.5mg / ml
[0092] Colcemid 15μg / ml
[0093] The stop solution formula of experimental group 3 is:
[0094] Ethidium Bromide 3mg / ml
[0095] Colcemid 12μg / ml
[0096] Set up a control group, in which:
[0097] The stop solution formula of the control group was:
[0098] Colcemid 12μg / ml
[0099] The steps for preparing bone marrow cell chromosome specimens ...
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