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Preparation of HeLa Nuclear Extracts

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In Vitro Transcription and Translation Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 37))

Abstract

Promoter-specific in vitro transcription by RNA polymerase II is now achievable with cell extracts prepared from a variety of animal and plant sources (1–3). For extracts prepared from mammalian cells, HeLa cells provide many advantages. Large quantities of homogeneous cells can be grown in spinner flasks at a cost below that of many other mammalian tissue-culture cells. A second advantage of HeLa extracts is the low level of RNases, proteases, and other nonspecific inhibitory activities. This results in a long shelf life (indefinitely at −70°C, <24 h at 4°C). The third advantage is that initiation factors from HeLa cells are well characterized, and many of them are available in recombinant form (4–10).

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Author information

Authors and Affiliations

  1. Center for Gene Regulation, Department of Molecular and Cell Biology, The Pennsylvania State University, University Park, PA

    B. Franklin Pugh

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  1. B. Franklin Pugh
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Editor information

Editors and Affiliations

  1. Monash University Clayton, Victoria, Australia

    Martin J. Tymms

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© 1995 Humana Press Inc.

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Pugh, B.F. (1995). Preparation of HeLa Nuclear Extracts. In: Tymms, M.J. (eds) In Vitro Transcription and Translation Protocols. Methods in Molecular Biology, vol 37. Humana Press. https://doi.org/10.1385/0-89603-288-4:349

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  • DOI: https://doi.org/10.1385/0-89603-288-4:349

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-288-0

  • Online ISBN: 978-1-59259-524-2

  • eBook Packages: Springer Protocols

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