Giemsa banding (GTC banding) procedure
Banding of chromosome with enzymes and stains is essential to identifying normal and abnormal chromosome structures.
- Gurrs 6.8 buffer tablets: (Cat. No. 10582013) one tablet (100 mL, 500 mL, 1 liter size) dissolved in distilled water
- 0.9% sodium chloride: 4.5 grams NaCl made up to volume of 500 mL distilled water
- Gurrs stain (R66) Giemsa:
- 3.0 mL Giemsa stain (Cat. No. 10092013) added to 48.5 Gurrs 6.8 buffer (unless otherwise stated on C of A)
- 1 mL acetone
- Mix in Coplin jar
- Trypsin 2.5%:
- 2.5 mL Trypsin (10X) (Cat. No. 15090046)
- 49.5 mL 0.9% NaCl
- Mix in Coplin jar
- Coplin jars or larger containers
- Coverslips (24 mm X 50 mm)
- 5 mL serological pipets (Cat. No. 170355)
- 1 mL serological pipets (Cat. No. 170353)
- 50 mL graduated cylinder
- 50°C warming oven
- Six Coplin jars are used for the banding sequence
- First jar—0.125% trypsin/0.9% NaCl mixture
- Second jar—0.9% NaCl for rinsing
- Third jar—0.9% NaCl for rinsing
- Fourth jar— Gurrs Giemsa stain (R66) mixed with Gurrs 6.8 buffer and acetone
- Fifth jar— Gurrs 6.8 buffer for rinsing
- Sixth jar— Gurrs 6.8 buffer for rinsing
- A slide is placed in the first Coplin jar containing the trypsin/NaCl mixture for a prescribed amount of time. This time may be as short as 10 sec or as long as 2 min, depending on the activity level of the trypsin being used.
- After the trypsin time has elapsed, the slide is removed and rinsed by sequential dipping into the 0.9% NaCl rinsing jars.
- The slide is then placed in the staining jar containing the Gurrs stain and buffer for 5 minutes. This time may vary somewhat, depending on the strength of the stain used.
- After the staining time has elapsed, the slide is removed from the jar and rinsed by sequential dipping into the two Gurrs buffer rinsing jars.
- The slide is removed from the last rinse and air dried and coverslipped with Cytoseal 60. It is allowed to dry in the oven (50°C) after which it is ready for metaphase scanning under the microscope.
- Breg, RW, Karyology and Chromosome Banding of Cultured Cells, Dec. 6-10, 1976 Lake Placid, NY.
- Sun, NC, Ehy, Chu, CC Chang (1976) Staining Method for Banding Patters Human Mitotic Chromosomes. MCN 14-26.