By Jose M Martinez-Zapater, Julio Salinas
This finished selection of present and crucial protocols comprises many simply reproducible equipment constructed to be used with Arabidopsis - a process for forthcoming primary questions in plant biology. The equipment diversity from the fundamentals of starting to be those vegetation to stylish gene cloning concepts and will, in lots of situations, even be utilized to different plant species with minor transformations. Sections on genetics, transformation and gene expression research which are specially priceless to scientists excited about mutant research or generating and interpreting transgenic vegetation.
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64, 83. 9. Browse, J. , and Somerville, C. (1986). , 81, 859. 10. Koornneef, M. and Hanhart, C. J. (1981). Arabidopsis Inf. , 18, 52. 11. Last, R. L. and Fink, G. R. (1988). Science, 240, 305. 12. Redei, G. P. and Acedo, G. (1976). In Cell genetics in higher plants, p. 39. Akad. Kiado, Budapest. 13. Conklin, P. , Williams, E. , and Last, R. L. (1996). Proc. Natl. Acad. Sci. USA, 93, 9970. 14. Relichova, J. (1976). Arabidopsis Inf. , 13, 25. 15. Hirono, Y. and Redei, G. P. (1963). Nature, 197, 1324.
B. Sterilization and sowing of seed 1. Wrap the seed in a 9 cm filter paper disc and place it in a 30 ml Universal container. 2. Add 20 ml of 70% (v/v) ethanol to the Universal containing the seeds and shake the container for 1 min. 3. Pour out the 70% (v/v) ethanol and replace it with 20 ml of the bleach solution. Shake the container gently for 10 min. 25 Mary Anderson and Fiona Wilson Protocol 5. Continued 4. Pour out the bleach solution and replace it with 20 ml of sterile distilled water. Shake the container for approx.
All waste solutions must be discarded into a beaker containing solid sodium thiosulfate. 4. Wash the seeds twice in distilled water for 15 min. 5. Open the 'Atmos' bag, pour the sodium sulfate inactivated waste into the sink, and wash down with plenty of water. Leave all beakers, etc. used in the procedure under running water for half an hour. 6. Transfer the seeds to 3MM paper and allow to dry overnight. 7. Dilute the seeds in dry sand to make distribution easier. Plant at a density of 1 cm-2.
Arabidopsis Protocols by Jose M Martinez-Zapater, Julio Salinas