A kind of preparation method of bone marrow chromosome g-band

Abstract

The invention discloses a manufacturing method for marrow chromosome G band, and the method comprises adding mankind lymphoma cell cultures into a marrow medium and adding a certain amount of ethidium bromide when cell culture is terminated. The effects comprise that (1) division phases are relatively more, time is saved and labor is saved; (2) the length of chromosome in division phases is relatively long, and the band is relatively clear; (3) the dispersity is relatively good and the analysis is facilitated; and (4) the detection rate of abnormal chromosome is relatively high and the diagnosis result is relatively reliable. The G band manufactured by employing the method has the advantages of saving time and saving labor when applied to marrowchromosome karyotyping, is relatively high in accuracy and has relatively wide application prospect.

Description

technical field

[0001] The invention belongs to the field of life science and biotechnology, and particularly relates to a method for making a G-band, which is used for bone marrow chromosome karyotype analysis. Background technique

[0002] 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.

[0003] Studies have found that human chromosome specimens are treated with reagents such as trypsin, Na0H, citrate or urea, and then stained with ...

Examples

Embodiment 1

[0051] Example 1 Preparation of bone marrow cell culture medium

[0052] Penicillin / streptomycin, fetal bovine serum, and human lymphoma cell cultures were added to the basal medium. The basal medium is RPMI1640 medium.

[0053] The consumption of described each added component is as follows:

[0054] Penicillin / Streptomycin 5-15ul / ml

[0055] Fetal bovine serum 60-140ul / ml

[0056] Human lymphoma cell culture 60-140ul / ml

[0057] Wherein, penicillin can be selected from 10000 U / ml, and streptomycin can be selected from 10000 μg / ml. In other embodiments, other concentrations of penicillin and streptomycin can be selected for the formulation of the medium.

[0058] Among them, human lymphoma cell culture can be obtained or prepared according to various existing methods. In this embodiment, it is prepared according to the following method: the cells are recovered and subcultured, and when the color of the subcultured cell culture medium turns yellow, collect Cells were cen...

Embodiment 2

[0069] Example 2 Bone Marrow Chromosome Extraction Kit

[0070] The bone marrow chromosome extraction kit includes liquid reagents composed of reagent 1, reagent 2, reagent 3 and reagent 4.

[0071] Wherein reagent 1 is a bone marrow cell culture medium, prepared according to the method described in Example 1, reagent 1 includes basal medium and each additional component, and the dosage of each additional component is:

[0072] Penicillin / Streptomycin 5-15ul / ml

[0073] Fetal bovine serum 60-140ul / ml

[0074] Human lymphoma cell culture 60-140ul / ml

[0075] Preferably, reagent 1 includes basal medium and each additional component, and the dosage of each additional component is:

[0076] Penicillin / Streptomycin 7-12ul / ml

[0077] Fetal bovine serum 80-120ul / ml

[0078] Human lymphoma cell culture 80-120ul / ml

[0079] More preferably, reagent 1 includes basal medium and each additional component, and the dosage of each additional component is:

[0080] Penicillin / Streptom...

Embodiment 3

[0108] Example 3 Bone marrow cell culture and preparation of chromosome samples

[0109] In this embodiment, bone marrow cells were cultured and chromosome preparations were prepared as follows. In other embodiments, other methods of culturing bone marrow cells and chromosome preparations can also be used. The method described in this embodiment is:

[0110] (1) Configure bone marrow culture medium: prepare according to the method in Example 1;

[0111] (2) Inoculation: inoculate the bone marrow cells into the medium described in step 1;

[0112] (3) Termination of cultivation;

[0113] (4) Harvest bone marrow cell culture for chromosome preparation;

[0114] (5) Chromosomal specimen preparation: Chromosomal specimens with G-banding were obtained after staining with dyes.

[0115] The medium and reagents required for bone marrow cell culture and chromosome extraction can be selected from the bone marrow chromosome extraction kit described in Example 2, or can be prepared ...

technical field

[0001] The invention belongs to the field of life science and biotechnology, and particularly relates to a method for making a G-band, which is used for bone marrow chromosome karyotype analysis. Background technique

[0002] 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.

[0003] Studies have found that human chromosome specimens are treated with reagents such as trypsin, Na0H, citrate or urea, and then stained with ...

Examples

Embodiment 1

[0051] Example 1 Preparation of bone marrow cell culture medium

[0052] Penicillin / streptomycin, fetal bovine serum, and human lymphoma cell cultures were added to the basal medium. The basal medium is RPMI1640 medium.

[0053] The consumption of described each added component is as follows:

[0054] Penicillin / Streptomycin 5-15ul / ml

[0055] Fetal bovine serum 60-140ul / ml

[0056] Human lymphoma cell culture 60-140ul / ml

[0057] Wherein, penicillin can be selected from 10000 U / ml, and streptomycin can be selected from 10000 μg / ml. In other embodiments, other concentrations of penicillin and streptomycin can be selected for the formulation of the medium.

[0058] Among them, human lymphoma cell culture can be obtained or prepared according to various existing methods. In this embodiment, it is prepared according to the following method: the cells are recovered and subcultured, and when the color of the subcultured cell culture medium turns yellow, collect Cells were cen...

Embodiment 2

[0069] Example 2 Bone Marrow Chromosome Extraction Kit

[0070] The bone marrow chromosome extraction kit includes liquid reagents composed of reagent 1, reagent 2, reagent 3 and reagent 4.

[0071] Wherein reagent 1 is a bone marrow cell culture medium, prepared according to the method described in Example 1, reagent 1 includes basal medium and each additional component, and the dosage of each additional component is:

[0072] Penicillin / Streptomycin 5-15ul / ml

[0073] Fetal bovine serum 60-140ul / ml

[0074] Human lymphoma cell culture 60-140ul / ml

[0075] Preferably, reagent 1 includes basal medium and each additional component, and the dosage of each additional component is:

[0076] Penicillin / Streptomycin 7-12ul / ml

[0077] Fetal bovine serum 80-120ul / ml

[0078] Human lymphoma cell culture 80-120ul / ml

[0079] More preferably, reagent 1 includes basal medium and each additional component, and the dosage of each additional component is:

[0080] Penicillin / Streptom...

Embodiment 3

[0108] Example 3 Bone marrow cell culture and preparation of chromosome samples

[0109] In this embodiment, bone marrow cells were cultured and chromosome preparations were prepared as follows. In other embodiments, other methods of culturing bone marrow cells and chromosome preparations can also be used. The method described in this embodiment is:

[0110] (1) Configure bone marrow culture medium: prepare according to the method in Example 1;

[0111] (2) Inoculation: inoculate the bone marrow cells into the medium described in step 1;

[0112] (3) Termination of cultivation;

[0113] (4) Harvest bone marrow cell culture for chromosome preparation;

[0114] (5) Chromosomal specimen preparation: Chromosomal specimens with G-banding were obtained after staining with dyes.

[0115] The medium and reagents required for bone marrow cell culture and chromosome extraction can be selected from the bone marrow chromosome extraction kit described in Example 2, or can be prepared ...

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