Stringency Buffer
| |
Stock Concentration |
Volume of Stock |
| 0.1xSSC |
20xSSC |
20ml |
| 10mM |
beta-mercaptoethanol |
3.5ml |
| 1mM EDTA |
250mM |
16.0ml |
| dH20 |
|
3960.5ml |
| Total Volume |
|
4000ml |
Dehydration Buffers:
| |
50% |
70% |
90% |
100% |
| 100% EtOH |
100ml |
140ml |
180ml |
200ml |
| 3M NH4Ac |
20ml |
20ml |
20ml |
-- |
| dH20 |
80ml |
40ml |
-- |
-- |
| Total Volumes |
200ml |
200ml |
200ml |
200ml |
Selected Bibliography
Wilcox, J.N., Gee, C.E., and Roberts, J.L. In situ cDNA:mRNA hybridization: Development of a technique to measure mRNA levels in individual cells. In: Methods in Enzymology, Vol 124, Neuroendocrine Peptides (P.M. Conn, ed.), Academic Press, pp510-533, 1986.
Wilcox, J.N., Smith, K.S., Williams, L.T., Schwartz, S., and Gordon, D. Platelet-derived growth factor mRNA detection in human atherosclerotic plaques by in situ hybridization. J. Clin. Invest. 82, 1134-1143, 1988.
Wilcox, J.N., Smith, K.M., Schwartz, S.M., Gordon, D. Localization of tissue factor in the normal vessel wall and in the atherosclerotic plaque. Proc Natl. Acad.Sci. 86, 2839-2843, 1989.
Melton, D.A., Krieg, P.A., Rebagliati, M.R., and Maniatis, T., Zinn, K., Green, M.R. Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter. Nucl. Acids Res. 12, 7035-7056, 1984.
Sources of Materials Used In Situ Hybridization
- United Dessicants, 6845 Westfield Ave., Pennsauken, NJ. 08110-1582 USA (609-662-6500)
- Humi-Cap dessicant capsules (No. 245-2)
- Baxter Scientific (now VWR Scientific)
- Nalgene utility boxes (No. L1995-4)
- Miles stain dishes (No. S7631-6)
- Miles stain racks (No. S7636)
- O.C.T. (No. M7148-4)
- Tissue culture chamber slides (No. T4135-4)
- Peel-a-Way tissue molds (No. M7275-3, No. M7275-2)
- VWR Scientific
- Micro slide boxes black (holds 25 each) (No. 48444-003)
- Slide grips for dipping in emulsion (No. 48440-002)
- Polysciences, Inc., 400 Valley Rd., Warrington, PA 18976 USA (800- 523-2575)
- Gills hematoxylin No. 2 (No. 4570)
- Alcoholic Eosin Y 1% (No. 17269)
- International Biotechnologies, Inc.(IBI), P.O. Box 9558, 275 Winchester Ave., New Haven, CT, 06535 USA (800-243-2555)
- Kodak NTB2 emulsion (No. 1654433)
- Whatman
- 3mm paper (No. 3030M917)
- DE81 filter paper (No. 3658M323)
- Fisher Scientific
- Autoimmerse heater (No. L1995-4)
- Superfrost/Plus Microscope Slides (No. 12-550-15)
- Promega Corp., 2800 Woods Hollow Rd., Madison, Wi 53711 USA (800-356-9526)
- Riboprobe Transcription buffer kits (ie. No. P2490)
- RNasin (No. N2511)
- Amersham Corp., 2636 South Clearbrook Dr., Arlington heights, IL 60005 USA (800-323-9750)
- 35S-UTP (for riboprobe transcriptions) (No. SJ1303 )
- 35S-dATP (low DTT concentration for tailing rxions) (No. SJ1334)
- Sigma Chemical, P.O. Box 14508, St. Louis, MO 63178 USA (1-800-325-3010)
- RNase A (No. R5125)
- Proteinase K (No. P4914)
- tRNA (No. R9001)
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