THEMMEN%HROEUR51@PUCC.PRINCETON.EDU (02/25/91)
Thomas Collet <collet@SCRIPPS.EDU> writes: >I am trying to clone immunoglobulins from hybridoma cells and >am wondering whether I am using the best possible restriction >enzymes. Does anyone have a compilation of the cutting >frequencies of restriction enzymes in immunoglobulin genes (or >maybe just mammalian genes in general ?) Or maybe a literature >reference ? >Thanks > >Thomas Thomas, The restriction site frequency can vary with the restriction enzyme's recognition pattern. In the New England Biolabs Catalogue you (88-89, p. 141 will find some info on the cutting frequency of some enzymes (NotI: (approx fragment size: 1000 kb) MluI, NruI, PvuI (300 kb) SfiI, RsRII (200 kb) SacII, BssHII, EagI, SmaI, SalI, AatII, XhoI, FspI, SnaBI, ClaI, AsuII (100 kb). As you can see all these enzymes have one or more CG's in their recognition seq. The above frequencies hold true for the mammalian genomes. Regards, Axel Themmen (Themmen@HROEUR51.bitnet)