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1、DIETHYL PYROCARBONATE Product Number D5758 Storage Temperature 2-8CCAS #: 1609-47-8 Synonyms: DEPC, DEP, Pyrocarbonic Acid Diethyl ester, Diethyl Oxydiformate, dicarbonic acid diethyl ester6Product Description Appearance: Clear colorless liquid Molecular formula: C6H10O5 Formula weight: 162.1 Densit
2、y: 1.12 g/mL Molarity: 6.9 M Refractive Index: 1.398 at 20CDiethyl Pyrocarbonate (DEPC) is sensitive to moisture and to pH; it decomposes to ethanol and carbon dioxide in aqueous solution. It decomposes at 155C. DEPC is also sensitive to ammonia, which causes decomposition to urethane, a possible ca
3、rcinogen.DEPC is effective as a nuclease inhibitor.2,3,4 (It reacts with many enzymes containing -NH, -SH or -OH groups in their active sites.8 Typically a 0.1% solution is used to inactive RNase (one milliliter of DEPC is added to 1 liter of water). The DEPC will not immediately dissolve, as eviden
4、ced by the appearance of globules. The mixture should be stirred until the globules disappear. The DEPC hydrolyzes over time to form ethanol and carbon dioxide. At this time the solution may be autoclaved to destroy the DEPC.DEPC has been used as a gentle esterifying agent and preservative.6 DEPC ac
5、ts as a condensing agent between lysine -amine groups and the carboxyl groups of aspartic or glutamic acid. It modifies purine residues in RNA by carboxyethylation of histidyl residues.4It is considered sufficient to autoclave 0.1% DEPC solutions for 15 minutes per liter.A method to assay for DEPC u
6、ses 5-thio-2- nitrobenzoate.4For general information and background, see references.Notes: 1. DEPC will dissolve some plastic pipets; therefore, glass should be used. It has been reported that it isincompatible with polycarbonate containers; it may react to form an insoluble solid. 2. If one does no
7、t wish to autoclave, the DEPC- treated water may be boiled (100C for 15 minutes). 3. Glassware is incubated with a DEPC solution for about 30 minutes and subsequently autoclaved to degrade the DEPCPreparation Instructions DEPC is tested for miscility in 95% ethanol; 1.6 mL in 4 mL ethanol gives a cl
8、ear, colorless solution. It is soluble in alcohols, esters, ketones and other hydrocarbons, but only soluble in water to about 0.1%.5 DEPC hydrolyzes in aqueous solution. In phosphate buffer at 25C, its half-life is 4 minutes at pH6, 9 minutes at pH 7. Hydrolysis is accelerated by TRIS buffer; at 25
9、C, its half-life was reported to be 1.25 minutes at pH 7.5, 0.37 minutes at pH 8.2.6Storage/Stability Diethyl pyrocarbonate is very sensitive to moisture, and as such, is packaged under argon to help reduce exposure to moisture. This product has a shelf-life of 2 years.If the product is exposed to e
10、ven traces of moisture, it does hydrolyze somewhat. The carbon dioxide which is formed is more soluble in DEPC at 0-4C than at room temperature, and as the product is brought to room temperature before opening, the DEPC at 0-4C can become supersaturated with respect to any dissolved gas. It is advis
11、ed that the user be familiar with the hazardous nature of DEPC, as described on the Material Safety Data Sheet (MSDS).After opening the bottle for the first time and each time thereafter, layer nitrogen or argon gas over the DEPC and store the closed bottle at 2-8C for optimal stability. It may be h
12、elpful to store the bottle inside a sealed plastic bag with desiccant, but with the bottle cap slightly loose. If the bag inflates at all, this is an indication of some degree of decomposition with possible pressure build-up. Once opened, the bottle should not be kept for more than 9 months.Referenc
13、es 1. Suppler data. 2. Williamson, R. et al., Biochemistry 10, 3014 (1971).3URGXFW,QIRUPDWLRQ3. Solymosy, F. et al., Eur. J. Biochem. 5, 520 (1968). 4. Sambrook, J., Fritsch, E.F., & Maniatis, T., Molecular Cloning: A Laboratory Manual, 2nd Ed. (1989) p. 7.3-7.4. 5. Berger, S.L. Analytical Biochem.,
14、 67, 428 (1975). 6. Merck Index, 12th Ed., p. 1375, #8182 (1996).7. Data For Biochemical Research, 3rd Ed., Dawson, R.M.C et al, ed., Oxford Press, 1984, p. 389. 8. Sigma Molecular Biology Department. 9. Ehrenberg, L. et al, Prog. Nucleic Acid Res. & Molec. Biol. 16, 189-263 (1976). “Diethyl Pyrocar
15、bonate in Nucleic Acid Research.feb 2/17/99Sigma brand products are sold through Sigma-Aldrich, Inc. Sigma-Aldrich, Inc. warrants that its products conform to the information contained in this and other Sigma-Aldrich publications. Purchaser must determine the suitability of the product(s) for their particular use. Additional terms and conditions may apply. Please see reverse side of the invoice or packing slip.
