By Steven M. Powell
Instructs investigators in key genetic, mobile, and molecular organic equipment of interpreting colorectal tumors. ideas and assays are defined intimately to permit researchers to scan on colon tumors. such a lot chapters are written by means of scientists who pioneered the tools provided. assurance contains gross microdissection of colon tumors, institution of phone traces and xenografted tumors, chromosomal analyses corresponding to FISH assays, mutational analyses of colon tumors and blood samples, gene expression analyses, and microsatellite instability trying out. incorporates a few colour photographs. For researchers at a variety of degrees, from graduate scholars to execs. Powell is affiliated with the collage of Virginia future health method.
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Additional resources for Colorectal Cancer: Methods and Protocols
Change the media again next day, leave overnight (see Note 3). 3. Harvesting 1. Add ethidium bromide (8 µg/mL) for one hour. 8 mg/mL) per 10 mL media. 2. After 1 h add colcemid (10 µg/mL) 100 µL to 10 mL flask and mix well by shaking or pipeting. 3. Transfer to 15 mL tube immediately. 4. Incubate at 37°C for 15 min. 5. Centrifuge for 5min at 1000 rpm. 6. Remove supernatant. 7. 075 M KCl/each tube drop by drop while tapping tubes. 8. Resuspend thoroughly using Pasteur pipet. 9. 075 M KCl to total 10 mL.
Et al. (1997) Somatic mutations of the MEN1 tumor suppressor gene in sporadic gastrinomas and insulinomas. Cancer Res. 57(21), 4682–4686. 22. , Vortmeyer, A. , Oldfield, E. , et al. (1997) Mutations of the MEN1 tumor suppressor gene in pituitary tumors. Cancer Res. 57(24), 5446–5451. 23. Lubensky, I. , Vortmeyer, A. , Libutti, S. , Choyke, P. , et al. (1998) Multiple neuroendocrine tumors of the pancreas in von Hippel- 50 24. 25. 26. 27. 28. 29. 30. 31. 32. 33. 34. 35. Pack and Zhuang Lindau Disease Patients: Histopathologic and Molecular Genetic Analysis.
Underdigested slides will have trim of the cytoplasm around cell nuclei and later give autofluorescence background. Digestion should proceed until nicely shaped clean nuclei show up. Overdigested specimens will display poor nuclei morphology, destructed chromatin. That slides should be discarded. 3. Change of the cell culture media each of 2 d before harvesting results in high mitotic index. 48 Pack and Zhuang 4. Optimum conditions for metaphase slide preparation should be determined first. If chromosomes overlap or retain the cytoplasm some modification can be used.