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EXTRACTION OF DNA FROM GOAT LIVER.pptx
- 1. EXTRACTION OF DNA FROM GOAT LIVER a M.Phool Badshah
- 2. Principle • DNA can be isolated from cells and tissues by high salt or phenol extraction. • Here, chromatin and proteins are dissociated by treatment with SDS. • The proteins are denatured by phenyl chloroform treatment. • It becomes insoluble and can be prepared by the following chemicals. Principle
- 3. Materials : • Ethanol • Saline • EDTA • pH-8.0 (0.9% NaCl and 0.01 M EDTA) • 10% SDS • 80% phenol • 95% ethanol Materials
- 4. A • 2.5 gm of liver tissue was homogenized in 25 mL of saline EDTA and transferred to a 250-mL conical flask. • To this, 2.5 mL of 10% SDS and 2.5 mL of 80% phenol was added and shaken well for 10 minutes. • The entire mixture was centrifuged at 10000 rpm for 10 minutes. The aqueous layer was separated out. Procedure
- 5. a • To the remaining material, (interphase + phenol phase), equal volume of chloroform, isoamyl alcohol (24 : 1) was added and shaken for 25 minutes. • It was again centrifuged at 10000 rpm for 10 minutes. • The aqueous phase was collected. • To the aqueous phase, equal volume of 80% phenol was added and again centrifuged at 10000 rpm for 10 minutes.
- 6. a • To the aqueous phase obtained, 95% ice cold ethanol was added and then the flask was shaken gently. • The DNA are precipitated as a white mass.
- 7. • By the addition of ice-cold ethanol, DNA strands were obtained as a white mass or turbidity. RESULTS A