CBG BANDING

I. Preparation

1. Get out 4 clean coplin jars.

2. Label them as follows:

• 0.2 N HCl
• 1% BaOH)2
• 2 X SSC
• Leishmans stain

3. Prepare 0.2 N HC1 as follows:

• 8 ml 1 N HCl
• 32 ml distilled water

4. Prepare 1% Ba(OH)2 as follows:

• 0.4 g Ba(OH)2
• 40 ml distilled H20

5. Prepare 2 X SSC as follows:

Pour 45-50 ml 2 X SSC into jar

6. Prepare Giemsa as follows:

• 1 ml Leishmans stain
• 3 ml Gurr buffer pH 6.8

II. Banding

1. Place slides in 0.2 N HCl for 1 hour.

2. Remove, rinse with distilled water.

3. Place slides in 1% Ba(OH)2 at 60° C for 15 seconds. Agitate the slide (BaOH2 will not go fully into solution.)

4. Remove, rinse with distilled water.

5. Place slides in 2 X SSC for 60° C for 1 hour.

6. Remove rinse with distilled water, allow to air dry.
(Do not use air jet.)

7. Stain slides in Leishmans for about 2-4 minutes.

8. Remove, rinse with tap water.

9. Air dry.

10. Ready for analysis

III. Reagents

Barium hydroxide--Biochem Stores

Gurr's Improved R66 Giemsa--Biomedical Specialties

Hydrochloric Acid 1N--Biochem Stores

Leishmans stain ---Sigma.

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