Recombinant DNA techniques: Storage and screening of cDNA libraries with large numbers of individual colonies from initial transformations
✍ Scribed by G. Vogeli; E. Horn; M. Laurent; P. Nath
- Publisher
- Elsevier Science
- Year
- 1985
- Tongue
- English
- Weight
- 287 KB
- Volume
- 151
- Category
- Article
- ISSN
- 0003-2697
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✦ Synopsis
A typical cDNA library with a large number of initial transformants, plated in soft agarose, can be stored and shipped in 12% glycerol at -70°C. To prepare the library for storage, the soft agarose layer is made into a paste and the agarose is removed by Sephadex G-25 filtration. This method of cDNA library storage does not alter the relative representation of the plasmids carried in the library. To achieve a very uniform distribution of colonies at high colony density, an aliquot of the cDNA library is diluted to 3000 to 10,000 colonies/ml. One milliliter of this suspension is evenly distributed on a nitrocellulose filter on an agar plate and air-dried. Filter copies are made and screened by published methods. o 1985 Academic PRS, IIIC.