mats@bio.embnet.se (Mats Sundvall) (10/28/90)
In article <9010231401.AA01888@genbank.bio.net>, MBY134@sysh.surrey.ac.uk writes: > Does anybody have any particular tips for doing inverse-PCR? > We've tried alterring magnesium conc., dilute ligations to encourage > intramolecular ligation, long extension times - but no product (dispite > direct PCR working on template) > Any ideas welcome. > > "Ever tried? Ever failed?. No matter > Try again. Fail again. Fail better! > Samual Beckett > > Johnjoe McFadden > University of Suttey, UkK > ACABCCCCCCCCCCCCCCCCCrreyBBBBBBBBAAAAAAAA, > CCCC A couple of new methods that is useful if you only know one end has been published. It all depends on your specific application if they are useful to you. Our experiance with inverse PCR are not the best. The methods we have used depend on ligation of a linker to the ends and amplifying only the ones with one internal primer. Variations one this is scheme is published as bubbel-PCR by Riley et al NAR Vol 18, No 10, page 2887. Another very similar method was published in a more recent NAR but I do not have this reference. Mats Sundvall Dep of Medical Genetics Uppsala University Sweden