By Michael Butler
This e-book is a wonderful source for college students and starting clinical reaseraches. it's a stable resource of simple equipment and methods for profitable laboratory phone tradition techniques. lots of the present cellphone tradition tools are coated yet this assurance isn't targeted. back, very good for rookies.
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Extra info for Animal Cell Culture and Technology (THE BASICS (Garland Science))
Transfer to a dry sterile tissue culture dish and chop into small pieces with sterile scissors. 8. 5 g glass beads (1 mm diameter) suspended in 1 ml PBS. 9. 2% (v/v) trypsin (1:1250) in Hank’s balanced salt solution and swirl the flask gently for 30 seconds. 10. Allow the larger fragments to sediment, then remove and discard the supernatant containing the erythrocytes. 11. Add a further 20 ml of trypsin, agitate for 30 seconds and transfer supernatant to a sterile centrifuge tube. Repeat this trypsin extraction ×5.
Layer 4 ml of diluted blood sample on to the Ficoll-Paque. 3. Centrifuge at 400 g for 30 minutes at room temperature. 4. Remove the upper layer (plasma) with a Pasteur pipette, leaving the lymphocyte layer undisturbed. 5. Transfer the lymphocyte layer into another centrifuge tube using a minimal volume. 6. Add three volumes of balanced salt solution. 7. Aspirate with a pipette to ensure adequate suspension of the cells in the liquid. 8. Centrifuge at 100 g for 10 min and discard the supernatant.
G. (1976) Stimulation of clonal growth of normal fibroblasts with substrata coated with basic polymers. J. Cell Biol. 71:727–734. A. G. (1981) The use of low-temperature subculturing and culture surfaces coated with basic polymers to reduce the requirement for serum macromolecules. G. J. et al. (eds) Requirements of Vertebrate Cells in vitro, pp. 118–130. Cambridge University Press, Cambridge. L. (1994) Sterilization. M. ) Basic Cell Animal cell culture and technology 54 Culture: A Practical Approach, pp.