PREPARATION OF SOMATIC CHROMOSOME FROM MAMMALION

SPECIMEN BY COLCHICINE- HYPOTONIC FLAME DRY GIEMSA

TECHNIQUE

●PROCEDURE IN SEQUENCE →

1) Inject colchicine to White Rat(Dosage -2mg/kg body weight)

2) Dissect of Specimen & collect bones including Femur
3) Flash two ends of bones with 0.075(M) Kcl
4) Collect bone marrow alongwith Kcl solution into Centrifuge tube
5) Incubate it at 37C0 for 20-30 mins
6) Homogenous suspension Centrifuge at 1000 rpm (10-15 mins)
At room temperature
7) Remove supernatant, pellets are taken
8) Add Fixative(Aceto alcohol-CH3OH : CH3COOH=3:1)
9) Suspension kept for 20 mins in chilled condition
10) Centrifuge as before & pellets are taken
11) Make suspension again with fixative(Aceto alcohol)
12) In 4 C0 kept it for overnight
13) 50% alcohol+clean slide in a beaker- Kept in chilled condition
for 1hrs
14) Centrifuge as before alongwith fixative & pellets are taken
15) Make suspension with fixative(Aceto alcohol), minimum
amount
16) With the help of Pipette sample put on to a clean slide & dry
by flame
17) Add Giemsa Stain (5-10 mins)
18) Rinse the slide in distilled water & ovserve under Microscope.