yablonsky@BIOVAX.RUTGERS.EDU ("Michael Yablonsky, Cook") (08/25/89)
Hello out there!!! Can someone advise me as to a strain of E. coli which will stably maintain repeated sequences? I'm cloning 25mers in DH5alpha and the clones are very unstable above 2-3 copies Any input will be appreciated!! Mike Yablonsky Yablonsky@biovax.bitnet Yablonsky@biovax.rut.edu
brutlag@NET.BIO.NET (Doug Brutlag) (08/25/89)
Mike, You might use the strains that we used to clone Drosophila satellite DNAs. We found that 60 to 100 copies of repeats 5 to 15 base pairs long were stable. The best references are: Lohe, A. R., & Brutlag, D. L. (1986). Multiplicity of satellite DNA sequences in Drosophila melanogaster.. Proc Natl Acad Sci U S A, 83(3), 696-700. Lohe, A. R., & Brutlag, D. L. (1987). Adjacent satellite DNA segments in Drosoph Lohe, A. R., & Brutlag, D. L. (1987). Adjacent satellite DNA segments in Drosophila:: structure of junctions.. J Mol Biol, 194(2), 171-9. Lohe, A. R., & Brutlag, D. L. (1987). Identical satellite DNA sequences in sibling species of Drosophila. J Mol Biol, 194(2), 161-70. However, your instability may also be sequence dependent. Doug Brutlag