Polynucleotide microarray including two or more groups of probe polynucleotides immobilized on substrate according to melting temperature and method for detecting target polynucleotides using the same

• US 7,338,764 B2
• Filed: 09/02/2004
• Issued: 03/04/2008
• Est. Priority Date: 12/03/2003
• Status: Expired due to Fees

• Current Assignee: Samsung Electronics Co. Ltd.
• Original Assignee: Samsung Electronics Co. Ltd.
• Inventors: Paek, Sanghyun
• Primary Examiner(s): WHISENANT, ETHAN C
• Application Number:
  US10/933,178
• Publication Number:
  US 20050123951A1
• Time in Patent Office: 1,279 Days
• Field of Search: 435/6, 536/23.1, 536/24.3
• US Class Current: 435/6.11
• CPC Class Codes: C12Q 1/6837 using probe arrays or probe chips C12Q1/6874 takes precedence
• CPC Class Codes: C12Q 2527/107 Temperature of melting, i.e. Tm
• CPC Class Codes: C12Q 2527/125 Specific component of sample, medium or buffer

Abstract

Provided is a polynucleotide microarray including two or more groups of spots on which probe polynucleotides are immobilized, wherein the probe polynucleotides of each group have melting temperatures (Tms) within a predetermined range of Tm between the probe polynucleotides and target polynucleotides. A method for detecting target polynucleotides using the polynucleotide microarray is also provided.

Claims

1. A polynucleotide microarray comprising

two or more groups of spots on which probe polynucleotides are immobilized,

wherein the probe polynucleotides are grouped according to a melting temperature (Tm) in a first hybridization solution between the probe polynucleotides and target polynucleotides, and

wherein probe polynucleotides of each group have melting temperatures (Tms) within a predetermined range;

and

a cover for each group of spots,

wherein the cover has an inlet and an outlet, and

wherein the cover and each group of spots can form an individual compartment.

2. The polynucleotide microarray of claim 1, wherein a difference in Tm between the spots of each group is 10°

C. or less.

3. The polynucleotide microarray of claim 1, wherein a difference in Tm between the spots of each group is 5°

C. or less.

4. A method for detecting target polynucleotides, which comprises

spotting samples containing target polynucleotides onto probe polynucleotides immobilized on each group of spots of a polynucleotide microarray, wherein the polynucleotide microarray comprises

two or more groups of spots on which probe polynucleotides are immobilized,

wherein the probe polynucleotides are grouped according to a melting temperature (Tm) in a first hybridization solution between the probe polynucleotides and target polynucleotides, and

wherein probe polynucleotides of each group have melting temperatures (Tms) in the first hybridization solution within a predetermined range;

treating each group of spots or the samples with a second hybridization solution,

wherein the second hybridization solution for a group of spots comprises a denaturant, a stabilizer, or a mixture thereof, and

wherein the second hybridization solution for a group of spots is selected such that for each probe polynucleotide of the group a difference between a Tm in the second hybridization solution between the probe polynucleotide and the target polynucleotides and a hybridization temperature (T) is within a predetermined range;

hybridizing the target polynucleotides to the probe polynucleotides immobilized on each group of spots at the hybridization temperature (T); and

detecting a result of the hybridization to detect the target polynucleotides.

5. The method of claim 4, wherein the target polynucleotides are labeled with a fluorescent marker and detecting the result of the hybridization uses a fluorescent light.

6. The method of claim 4, wherein the denaturant is selected from the group consisting of formamide, urea, guanidine chloride, dimethylformamide (DMF), and a mixture thereof.

7. The method of claim 4, wherein the stabilizer is a salt selected from the group consisting of Na⁺, K⁺, Ca²⁺, and Mg²⁺.

8. The method of claim 4, wherein for each group of spots the difference between the Tms and the hybridization temperature (T) is in a range of 0 to 5°

C.

9. The method of claim 4, wherein for each group of spots the deviation of the Tms and the hybridization temperature (T) is in a range of 0 to 3°

C.

10. The method of claim 4, wherein each group of spots is covered with a cover,

wherein the cover has an inlet and an outlet, and

wherein the cover and the group of spots form a compartment.