Duplex DNA is isolated from bacteriophage lambda (cI857ind 1 Sam 7). The bacteriophage is grown in an E. coli host which is deficient in adenine methylase (dam-). Therefore, those deoxyadenosine positions which are methylated in a wild type E. coli host are not methylated in the dam- host. This DNA gives complete digests with restriction endonucleases that are sensitive to dam methylation. Lambda DNA (dam-) is 48,502 base pairs in length.
Lambda map
Source
The phage is isolated from a heat-inducible E. coli(dam-) lambda lysogen. The DNA is isolated from purified phage by phenol extraction and dialyzed against 10 mM Tris-HCl (pH 8.0)and 1 mM EDTA.
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The advantage of this DNA is that it gives complete digests with those restriction endonucleases (BclI, MboI, DpnII, etc.) whose recognition sites are partially modified by most E. coli strains.
References
Daniels, D.L. et al. (1983). Appendix II: Complete Annotated Lambda Sequence. R.W. Hendrix, J.W. Roberts, F.W. Stahl and R.A. Weisberg(Ed.), Lambda-II. 519-676. New York: Cold Spring Harbor Laboratory Press.
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