Vectors and Bacterial Strains

Bacteriophage A. arms

If making a large number of libraries, it is much less expensive to prepare arms by agarose gel electrophoresis or by sucrose density gradients (please see Chapter 2, Protocol 16) than to purchase them. For occasional users, dephosphorylated arms for some vectors (e.g., ?^gt10, A.Fix, and T^ZipLox) are available from Stratagene, Life Technologies, and other commercial suppliers. It is important to carry out a series of pilot reactions to check that the arms can be ligated to foreign DNA and packaged efficiently into infectious bacteriophage A, particles. DNAs for these controls are usually included with commercial preparations of vector arms.

E. coli strain, freshly prepared overnight cultures

Use strains C600 (BNN93) for growth and BNN102 (C600h\\k) for screening ofcDNA libraries constructed in bacteriophage ?^.gt10. Use strain Y1090hs6R for growth and screening ofcDNA libraries constructed in bacteriophage Agf?1. Use strain BB4 for growth and screening ofcDNA libraries constructed in "K-ZAP and 'kZAPU or strain XL1-Blue forXZAPIi. (Strain XL1-Blue supports vigorous growth ofXZAPil but not of XZAP.) ForXZipLox, use strain Y1090(ZL). Packaging extracts for bacteriophage A,

Packaging extracts are difficult to make and should therefore be purchased commercially. Several commercial suppliers provide packaging kits with unique features.

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