Methods for genotype screening using magnetic particles
This invention relates to a method of an assay of genomic nucleic acid. Additionally, this invention relates to various methods to detect or screen for designated genetic sequences or portion thereof derived from a biological sample. More particularly, this invention relates to the use of magnetically responsive magnetic particles that function to reversibly bind genomic nucleic acid. A portion of the purified genomic nucleic acid is detected in solution by a variety of methods.
1. A method for detecting a nucleic acid sequence comprising:
treating a mammalian sample to obtain cellular debris including genomic nucleic acid;
mixing said cellular debris and a plurality of magnetically-responsive particles under conditions to facilitate reversible immobilization of said genomic nucleic acid to said plurality of magnetically-responsive particles, wherein said magnetically-responsive particles have a coating of glass microfiber;
removing the portion of said cellular debris not reversibly immobilized to any of said magnetically-responsive particles;
adding to said magnetic particles under hybridizing conditions nucleic acid probes capable of hybridizing to a portion of said genomic nucleic acid resulting in hybridized nucleic acid probes and unhybridized nucleic acid probes;
removing said unhybridized nucleic acid probes; and
detecting said hybridized nucleic acid probes.
2. The method of claim 1 further comprising:
eluting said genomic nucleic acid from said plurality of magnetically-responsive particles.
3. The method of claim 1 wherein said nucleic acid probes are ligated to a dendrimer.
4. The method of claim 3 further comprising the steps of:
releasing said dendrimer from said nucleic acid probes; and
detecting said dendrimer in solution.
5. The method of claim 1 wherein said nucleic acid probes react with elements on a dendrimer.
6. The method of claim 1 wherein said hybridized nucleic acid probes are disassociated from said genomic nucleic acid prior to detecting in solution.
7. The method of claim 1 wherein said mammalian sample is embryonic.
8. An assay for a nucleic acid sequence comprising:
treating a mammalian sample to obtain cellular debris including genomic nucleic acid;
mixing said cellular debris and a plurality of magnetically-responsive particles under conditions that facilitate reversible immobilization of said genomic nucleic acid to said plurality of magnetically-responsive particles, wherein said magnetically-responsive particles have a coating of glass microfiber;
removing the portion of said cellular debris not reversibly immobilized to any of said magnetically-responsive particles;
eluting said nucleic acid from said plurality of magnetically-responsive particles;
adding nucleic acid probes capable of hybridizing to a portion of said genomic nucleic acid under hybridizing conditions to form genomic nucleic acid-nucleic acid probe duplexes while in solution;
adding a plurality of magnetically-responsive particles to bind the genomic nucleic acid-nucleic acid probe duplexes while in solution;
removing unbound nucleic acid probes from said solution;
eluting said genomic nucleic acid-nucleic acid probe from said plurality of magnetically-responsive particles; and
detecting hybridized nucleic acid probes in said solution.
9. The assay of claim 8 wherein said nucleic acid probes are ligated to a dendrimer.
10. The assay of claim 9 wherein said nucleic acid probes are reactive with elements on said dendrimer.
11. The method of claim 9 further comprising the steps of:
releasing said dendrimer from said nucleic acid probes; and detecting said dendrimer in solution.
12. The assay of claim 8 wherein said hybridized nucleic acid probes are disassociated from said genomic nucleic acid prior to detecting in solution.
13. The assay of claim 8 wherein said mammalian sample is embryonic.