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Monday, 26 November 2007

genetic engineering (10)DNA preparation for cloning

DNa preparation from cell
keep Easy, Feasible, Simple

Animal cells:
Liver, spleen,kidney, blood, cell culture
Liver: fast 24 hr to decrease glycogen
Spleen: good source, large amount

fresh tissue or frozen tissue (-80 can keep for 1 year)

Principles see isolation DNA protocol
1. Cell lysis by detergent: SDS, sarkosyl, proteinase K
2. Extract protein and cell debris with phenol
3. Pecipitate DNA with ethanol, isopropanol
3. Purification by high g centrifugation in CsCl, ethidium bromide
Electrophoresis to measure size , quantitate with OD method

Plant cells : ask botanist I don't know much about these.

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