FRED@blekul13.bitnet (fred) (06/28/90)
Admitting the superiority of the people listed below is not easy. But then again we never really believed a world record was in the making here. I do appreciate the time you took to respond. Maybe we can build a "usefull-thing-to-know-but-kept-silent-for-unknown-reasons"-kind of mailbox here. List of responses: ========================================================================= Date: Wed, 27 Jun 90 15:33:31 -0700 From: "Marc D. Perry" <mdperry@boulder.Colorado.EDU> Subject: Re: king-size-cloning To: FRED@BLEKUL13.BITNET Organization: MCD Biology, University of Colorado, Boulder Fred, Many of the early ColE1/pMB9 derived vectors (from which pUC's are derived) were 10 kb, and genomic libraries constructed in them had avg. insert sizes of 10 kb (10 + 10 = 20). So I wouldn't call Guinness just yet. Marc P. --------------------------------------------------------- Marc D. Perry (303) 492 8258 MCD Biology, Univ. of Colorado, Boulder, CO 80309-0347 internet: mdperry@boulder.colorado.edu -- Marc D. Perry (303) 492 8258 MCD Biology, Univ. of Colorado, Boulder, CO 80309-0347 internet: mdperry@boulder.colorado.edu ========================================================================= Date: Wed, 27 Jun 90 10:52 EDT From: Department of Molecular Karma <YABLONSKY@MBCL.BITNET> Subject: Re: KING-SIZE-CLONING To: FRED@BLEKUL13.BITNET Minor league....but keep trying!!! ========================================================================= Date: Tue, 26 Jun 90 13:25 EST From: SCOLNIPA%ESVAX@dupont.com Subject: RE: king-size-cloning To: FRED@blekul13.BITNET X-Vms-To: ESPRNT::IN%"FRED%blekul13.bitnet@genbank.bio.NET" No way. We have several plant clones in pUC that are 15 kb or bigger. ========================================================================= Date: Tue, 26 Jun 90 10:58:21 EST To: fred@blekul13.bitnet Subject: king-size-cloning I was not too surprised. I had cloned several 10 to 16 kb inserts in pUC plasmids. In addition cosmids (derivatives of pUC plasmids with cos site) take up 40 kb inserts and stably replicate as plasmids. K. Bhat ========================================================================= Date: Tue, 26 JUN 90 17:39:05 GMT From: PHILLIPSA@ LARS.AFRC.AC.UK To: METHODS@ DLGM.DARESBURY.AC.UK Re: KING-SIZE-CLONING Fred, My record is a 21.9 kbp fragment of tomato chloroplast DNA in pUC8 (A. Phillips, Curr. Genet (1985) 10:147-152. Raise you 13kbp? Andy ========================================================================= Date: Tue, 26 Jun 90 07:14:18 EDT To: FRED@blekul13.BITNET Subject: Re: king-size-cloning Fred, I don't find subcloning a 9 kb fragment into pUC unusual. We have inserted 8 and 10 kb fragments before. douglas prasher woods hole oceanographic inst =============================================================================== =============================================================================== FRED at BLEKUL13. Acknowledge-To: <FRED@BLEKUL13>
rabani@sdbio2.ucsd.edu (Ely Rabani/D Smith) (07/02/90)
In article <9006261058.AA25702@genbank.bio.net> FRED@blekul13.bitnet (fred) writes: >The fun thing is that she has managed to clone a 9 kB SacI fragment >into the 2.7 kB pUC vector and that the thing is behaving completely >normal in our E.Coli as if it was not 11.7 kB big. Our question now: >is this the world record or are we too optimistic? Unless someone >says that these vectors can do more than expected we want to believe >that this is an exceptionally big insert for pUC. Is this Don't really know about pUC, but supposedly inserts >40kb (!!!!!) have been successfully cloned into M13...saw this cited someplace in Methods in Enzymology, think the reference was Michael Bevan. ER