@article{8a55139690b4464b974b1973d10b1151,
title = "PIG-B: A homemade monophasic cocktail for the extraction of RNA",
abstract = "An inexpensive monophasic reagent has been developed for the extraction of total RNA from cells or tissues. The main ingredients of the reagent are Phenol, Isoamyl alochol, Guanidinium isothiocyanate, and Beta-mercaptoethanol (PIG-B). The quality and yield of RNA obtained by this reagent is at par with that obtained by TRIzol, an expensive but widely used monophasic reagent available commercially. The complete composition and method of preparation of PIG-B is provided to aid preparation of the reagent in the laboratory.",
keywords = "RNA extraction, RT-PCR amplification, Reagent",
author = "Kristy Weber and Bolander, {Mark E.} and Gobinda Sarkar",
note = "Funding Information: 1. TRlzol was used as a reference reagent because the basic composition of TRlzol appears to be most comparable to PIG-B. 2. RNA was extracted separately with TRIzol and PIG-B from each tissue sample or cell culture. 3. The basic RNA extraction protocol is the same for both TRIzol and PIG-B. 4. Unlike TRIzol, the composition of PIG-B includes 8-Hydroxyquinoline. This is a partial inhibitor of RNase and an antioxidant, thus a reagent containing this compound should pro-vide better protection against RNases (I) and also improve the stability of phenol which is oxidized by air. In addition, 8-Hydroxyquino-line is a weak chelator of metal ions (6), which should also aid in inactivating any contaminat-ing nucleases. 5. PIG-B also contains isoamyl alcohol, which acts as an antifoaming agent during extraction of nucleic acids and thus facilitates the separa-tion of organic and aqueous phases (1). 6. Because of these differences in the chemi-cal composition between TRIzol and PIG-B, the latter may provide better protection against nucleases and result in a better RNA yield. 7. Our preliminary results indicate that the quantity of RNA recovered from the rabbit and human tissues was greater using the PIG-B reagent as compared to the commercial preparation. 8. Because of the presence of isoamyl alcohol, PIG-B may have longer shelf-life than TRIzol. 9. We have not investigated if PIG-B can be used for extracting DNA. Acknowledgment We thank Drs. Arunik Sanyal and Shawn O'Driscoll for the rabbit tissues used in this investigation. This work was supported by an NIH grant to M.E.B (2ROIAR41917-04), the Mayo Foundation, and the Mayo Cancer Center.",
year = "1998",
doi = "10.1007/BF02752699",
language = "English (US)",
volume = "9",
pages = "73--77",
journal = "Applied Biochemistry and Biotechnology - Part B Molecular Biotechnology",
issn = "1073-6085",
publisher = "Humana Press",
number = "1",
}