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        Preparation of HeLa Nuclear Extracts

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        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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