All Submissions | J. Saba Submissions | Physics Site Links | Home Page |
| Email: James Saba |
GSJ:Received May 18, 2005:
http://wbabin.net/saba/saba41.htm
Whole Genome Amplification Utilizing Nick Displacement Amplification
James Saba
It becoming increasing clear that whole genome (global) amplification is becoming a valuable technology, and their are various ways which it is achieved (1,2).
Herein is disclosed yet another method, and it is based on previously described nick displacement amplification (NDA) utilizing hybrid DNA/RNA primers wherein the 3' nucleotides are, after functioning in polymerase priming, are sites of RNaseH nicking (3,4).
Figure 1 is an extremely simplified cartoon of what is basically happening. Two other versions found in the original disclosure (1) are reproduced in the
appendix.
Notice the primers could be designed to prevent their displacement and block polymerization.
Conceivably, only one NDA primer sequence would be sufficient, yet multiple NDA primers of different sequence could be used. So to, a combination of different types of primers is possible
In the broadest sense of the invention any NDA primer, such as those nicked by nicking enzymes (5), could be utilized.
The ability to limit the amount of primer used, and more precisely control sizes of the amplification products, may provide distinct advantages to current methods of whole genome amplification.
This invention, as most of my inventions described in this journal, is at the conceptual stage and a patent is anticipated. However, it is hoped that those with laboratories will investigate its full potential.
Claims
2) The process of claim 1 wherein the NDA primers are hybrid primers comprising 3' ribonucleotides, and nicked by an RNaseH.
2)
The use of whole genome amplification in the study
of human disease.
Hughes, et al Prog Biophys Mol
Biol. 2005 May;88(1):173-89
3)
An Isothermal Nucleic Acid Amplification (Nick
Displacement Amplification).
Saba, JA 2004 May 16
4) Method for amplifying nucleic acid sequence.
Mukai, et al US Patent Appl 20030073081 April 17,
2003
5)
Isothermal reactions for the amplification of
oligonucleotides.
Van Ness J, et al Proc Natl Acad Sci
U S A. 2003 Apr 15;100(8):4504-9
6) Parallel genotyping of over 10,000 SNPs using a
one-primer assay on a high-density oligonucleotide
array.
Matsuzaki, et al Genome Res. 2004
Mar;14(3):414-25
