Length polymorphisms in (dC-dA).sub.n .multidot.(dG-dT).sub.n sequences

• US 5,075,217 A
• Filed: 04/21/1989
• Issued: 12/24/1991
• Est. Priority Date: 04/21/1989
• Status: Expired due to Term

• Current Assignee: Marshfield Medical Research Foundation
• Original Assignee: Marshfield Clinic
• Inventors: Weber, James L.
• Primary Examiner(s): Wax, Robert A.
• Assistant Examiner(s): Fleisher, Mindy B.
• Application Number:
  US07/341,562
• Time in Patent Office: 977 Days
• Field of Search: 435/6, 536/27, 935/77, 935/78
• US Class Current: 435/6.11
• CPC Class Codes: C12Q 1/6858 Allele-specific amplification
• CPC Class Codes: C12Q 1/6876 Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
• CPC Class Codes: C12Q 2600/156 Polymorphic or mutational markers

Abstract

The present invention is directed to a product and process for characterizing the human genome utilizing polymorphic DNA fragments containing (dC-dA)_n ·(dG-dT)_n. The process comprises identifying polymorphic DNA fragments containing the above tandem repeat sequence followed by characterizing admixtures of genomic material by amplification of fragments containing the tandem repeat sequence using substantially unique and non-polymorphic primer sequences and appropriate separation of the so amplified fragments.

Claims

1. A method of analyzing or typing polymorphic DNA fragments which contain (dC-dA)_n.(dG-dT)_n repeats which are specific to particular loci in the genome comprising:

amplifying at least one polymorphic DNA fragment containing at least one (dC-dA)_n.(dG-dT)_n segment using the polymerase chain reaction with a DNA sample which includes the said DNA fragment as a template, and with two oligonucleotide primers sufficiently complementary to non-repeated sequences at the ends of said fragment within said sample to hybridize therewith, and separating and characterizing amplified fragments on the basis of size.

2. The method of claim 1 further comprising labeling or marking said polymorphic DNA fragments.

3. The method of claim 1 wherein said primers are obtained by direct synthesis.

4. The method of claim 1 wherein said primers are end-labeled.

5. The method of claim 1 wherein more than one DNA fragment containing at least one segment of (dC-dA)_n.(dG-dT)_n is amplified and characterized simultaneously.

6. The method of claim 1 used to establish the identity, pedigree, or relationship of an individual comprising comparison of said amplified fragments with amplified fragments from the DNA of one or more control individuals wherein the control DNA samples have been similarly amplified and characterized.

7. The method of claim 1 wherein the amplified fragments are separated and characterized by gel electrophoresis.

8. The method of claim 1 wherein the DNA sample is genomic DNA.

9. The method of claim 1 wherein the DNA sample is cloned DNA.