Chromosome G Banding Technology Protocol
GUIDELINE
After treatment with physical and chemical factors, the chromosomes are then differentiated and stained with dyes, so that each chromosome appears on the light and dark, or different shades of banding technique is called banding technique. The number, location, width, and coloring depth of the bands are relatively stable, so each chromosome has a fixed banding pattern, known as a banding pattern. Chromosome banding is an important basis for identifying chromosomes.
METHODS
- The prepared chromosome specimens are baked in an oven at 72-75°C for 3 h.
- Put into the pre-warmed to 37°C. 0.025% trypsin solution (pH = 7.2-7.4) digestion for about 1 min, each time the action time is not the same. You can try one slice first, and then adjust the trypsin action time according to the effect of banding.
- Staining in Giemsa staining solution for 10 min, rinsed with tap water, dried, and ready to examine.
- Examine the banded specimen under the high magnification eyepiece of the microscope. If there are clear bands of different shades on the chromosomes, the specimen is considered to be acceptable.
Creative Bioarray Relevant Recommendations
- G-banding is a technique used in cytogenetics to produce a visible karyotype by staining condensed chromosomes. Creative Bioarray has performed many karyotyping (G-banded) services for cell line authentication and some specific research.
NOTES
- Trypsin concentration and processing time vary with temperature. The general rule is that the longer the specimen has been stored, the longer it will be processed in trypsin. Specimens that are too fresh will have fuzzy chromosomes. Very old specimens often result in blotchy chromosomes.
- If the cells are purplish-blue, the trypsin has not been working long enough, and if the cells are peach-colored, it is working just long enough.
