RE (MspI) Digestion of PCR Products

Martin Leach leach at
Sun Apr 16 20:06:35 EST 1995


I remember looking at polymorphic sequences using ASO (allele specific 
oligonucleotide) hybridization. We were using a TMAC (tetra-methyl ammonium 
chloride) style hybridization / washing procedure (i believe maniatis now has an 
entry on this).

Using this technique we could detect homo/hetero zygotes on a slot blot of PCR 
products,  for polymorphishms in the prion gene..

for more info see: (number or 2 or 5 - sorry don't have the paper handy..if these 
don't mention it they should have a reference to it..I think J. Collinge published a 
paper later on in Nature looking at sporadic CJD and polymorphic sequences..)

2.M. Poulter, H.F. Baker, C.D. Frith, M. Leach et. al. (1992). Inherited Prion
              Disease With 144 Base Pair Gene Insertion I: Genealogical and 
Molecular Studies. Brain 115: 675-685.

           3.F. Owen, M. Poulter, J. Collinge, M. Leach et. al. (1991). Insertions 
in the Prion
              Protein Gene in Atypical Dementias. Experimental Neurology 112: 

           5.J. Collinge, F. Owen, M. Poulter, M. Leach et. al. (1990). Prion 
              Without Characteristic Pathology. Lancet 336: 7-9


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