Claims
- 1. A method for preparing a heteroduplex-enriched nucleic acid sample from an organism carrying heterozyygous mutations, comprising the steps of:
- (a) denaturing substantially all of the double-stranded nucleic acid present in a nucleic acid sample from an organism;
- (b) allowing the nucleic acid to anneal the under conditions which permit formation of heteroduplexes and homoduplexes; and
- (c) removing homoduplexes from the annealed sample wherein the removal of said homoduplexes is effected using immobilized mismatch binding proteins (MBP), thereby retaining heteroduplexes in said sample.
- 2. The method according to claim 1, wherein said organism is a mouse.
- 3. The method according to claim 1, further comprising prior to step (a) the step of fragmenting the nucleic acid in the sample.
- 4. The method according to claim 1, wherein the removal of homoduplexes is effected using immobilized MutS protein.
- 5. The method according to claim 1, further comprising prior to step (a) the steps of: exposing the germ cells of a first parent organism to mutagenic conditions; and mating the first parent organism to a second parent organism to produce an offspring organism carrying heterozygous mutations.
- 6. A sample of nucleic acid heteroduplexes obtained by the method of claim 5, wherein the frequency of mutation carried in the offspring organism is higher than that of spontaneous mutation.
- 7. A method for screening a population of organisms carrying heterozygous mutations in order to identify an organism of the population which carries a mutation in a gene of interest, comprising the steps of
- i. preparing a plurality of samples of nucleic acid from a corresponding plurality of organisms of a population according to the method of claim 1 to produce a plurality of samples, and
- ii. contacting the plurality of samples with a probe specific for a gene of interest so as to identify a mutation in a gene of interest in an organism containing a mutation.
- 8. The method according to claim 7, wherein said plurality of organisms comprises 100 or more organisms.
- 9. The method according to claim 7, wherein said samples are affixed at defined positions to a solid support prior to said contacting step.
- 10. The method of claim 3, wherein the fragments of the nucleic acid in the sample are 200 to 400 base pairs in length.
Parent Case Info
This application claims the benefit of U.S. Provisional Application No. 60/066,206, filed Nov. 19, 1997.
Non-Patent Literature Citations (1)
Entry |
Casna et al., Nuc. Acids Res., vol. 14, 1986, 7285-7303. |