Selection of simple repeats (STR`s): A rapid technique using superparamagnetic beads
- Univ. of California, Los Angeles, CA (United States); and others
Great interest has been generated in the past few years for acquiring repeat sequences like di-, tri- and tetra-nucleotide repeats for their wide use as polymorphic markers. In this protocol we describe using an oligonucleotide (CA){sub 21} as a primer to extend and biotinylate the DNA of interest using Taq DNA polymerase. Streptavidin-coated superparamagnetic beads were then used to select the biotin-labeled DNA. The selected DNA can then be cloned quickly. The usefulness of the technique is its improved specificity achieved by controlling conditions for varying lengths and nucleotide composition of repeats. The other advantage is that, for this method, many different oligonucleotides can be combined in the same reaction and because Taq DNA polymerase is used, a very wide spectrum of annealing temperatures can be applied. These experiments were performed with a 9600 Perkin Elmer PCR machine. Reliable and reproducible results were obtained. The protocol circumvents the radioactive hybridization steps and can be completed in a short period of time.
- OSTI ID:
- 134133
- Report Number(s):
- CONF-941009-; ISSN 0002-9297; TRN: 95:005313-0869
- Journal Information:
- American Journal of Human Genetics, Vol. 55, Issue Suppl.3; Conference: 44. annual meeting of the American Society of Human Genetics, Montreal (Canada), 18-22 Oct 1994; Other Information: PBD: Sep 1994
- Country of Publication:
- United States
- Language:
- English
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