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

Active Publication Date: 2014-04-23
南京艾迪康医学检验所有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the interference of a large number of fat particles in the bone marrow, the cell cycle of various cell lines in the bone marrow is not fixed, not uniform, and it is diffi

Method used

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  • Preparation method of marrow G-band for karyotype analysis
  • Preparation method of marrow G-band for karyotype analysis
  • Preparation method of marrow G-band for karyotype analysis

Examples

Experimental program
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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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Abstract

The invention discloses a preparation method of a marrow G-band for karyotype analysis in the medical examination field. The method includes adding ethidium bromide and demecolcine into a medium at the end of culture. The adding of a certain amount of ethidium bromide (EB) at the end of cell culture can make the chromosome in a split phase longer and the band patterns clearer. With the advantages of time saving, labor saving, and higher accuracy, the marrow G-band has more extensive application prospects in the field of medical examination.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for preparing bone marrow G-bands for chromosome karyotype analysis in the field of medical testing. [0002] Background technique [0003] G-banding is called G-banding technology because the chromosomes are mainly stained with Giemsa dye, and the banding patterns displayed are distributed on the entire chromosome. People use various methods and different dyes to process chromosome samples to make each chromosome appear light and dark, or the technology of different shades of banding is called banding technique (banding technique). Since the 1970s, the banding technology has been greatly developed, and among the many banding technologies (Q band, G band, C band, R band, T band), the G band is currently widely used. belt type. [0004] Studies have found that human chromosome specimens are treated with reagents such as trypsin, Na0H, citrate or urea, and then ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68G01N1/30
CPCG01N1/2813G01N1/30
Inventor 程建兵夏成青陈红梅郭福晓生帅
Owner 南京艾迪康医学检验所有限公司
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