Detection oligomer and method for controlling quality of biochip using detection oligomer

• US 8,313,905 B2
• Filed: 01/23/2009
• Issued: 11/20/2012
• Est. Priority Date: 01/24/2008
• Status: Expired due to Fees

• Current Assignee: Samsung Electronics Co. Ltd.
• Original Assignee: Samsung Electronics Co. Ltd.
• Inventors: Kim, Byung-chul, Jung, Sun-Ok, Cho, Seung-Hei, Rhee, Joo-Won
• Primary Examiner(s): KAPUSHOC, STEPHEN THOMAS
• Application Number:
  US12/358,530
• Publication Number:
  US 20090239770A1
• Time in Patent Office: 1,397 Days
• Field of Search: None
• US Class Current: 435/6.1
• CPC Class Codes: C12Q 1/6837 using probe arrays or probe chips C12Q1/6874 takes precedence
• CPC Class Codes: C12Q 2521/501 Ligase
• CPC Class Codes: C12Q 2525/131 incorporating a restriction site
• CPC Class Codes: C12Q 2525/301 Hairpin oligonucleotides

Abstract

A detection oligomer and method for controlling the quality of a biochip using the detection oligomer are provided. The detection oligomer includes a template having a first and second end and having a sequence that is complementary to a specific sequence of an oligomer, and a hairpin connected to the first end of the template.

Claims

1. A method of assessing biochip quality, the method comprising:

providing a biochip comprising at least first full length polynucleotide probes and second truncated polynucleotide probes, each having first ends that are coupled to an immobilization layer, the first full length polynucleotide probes having a different number of nucleotide monomers than the second truncated polynucleotide probes;

hybridizing Quality Control (QC) probes to the first full length polynucleotide probes and second truncated polynucleotide probes, each QC probe comprising;

a template portion having a nucleotide sequence complementary to a nucleotide sequence of the first full length polynucleotide probes and second truncated polynucleotide probes; and

a hairpin connected to a first end of the template portion;

exposing the biochip to conditions suitable for ligation of polynucleotides;

washing the biochip to de-hybridize the QC probes from the first full length polynucleotide probes and second truncated polynucleotide probes, thereby removing un-ligated QC probes;

detecting a ratio of the amount of the first full length polynucleotide probes that are ligated to the QC probes to the amount of the second truncated polynucleotide probes that are ligated to the QC probes to assess biochip quality, wherein the quality of the biochip is a measure of the relative amounts of first full length polynucleotide probes and second truncated polynucleotide probes.

2. The method of claim 1, further comprising:

phosphorylating second ends of the first full length polynucleotide probes and second truncated polynucleotide probes before the hybridizing.

3. The method of claim 1, wherein:

the hairpin comprises a stem comprising complementary first and second single strands and a loop, the first single strand having a first end that is coupled with the loop and a second end that is coupled with a first end of the template portion, and the second single strand having a second end that is coupled with the loop and a first end that is ligated to the second end of the first full length polynucleotide probe during the exposing the biochip to conditions suitable for ligation.

4. The method of claim 1, wherein:

the hairpin comprises a stem having complementary first and second single strands and a loop; and

after the hybridizing, first ends of the second strands of the QC probes hybridized to the second truncate polynucleotide probes are spaced apart from the second ends of the second truncated polynucleotide probes.

5. The method of claim 1, further comprising:

after the washing, re-hybridizing the full length polynucleotide probes to the template portion of the ligated QC probes to form double stranded structures;

intercalating a label into the double stranded structures; and

detecting the label.

6. The method of claim 1, wherein:

the QC probes each further comprise a label coupled to one of the template portion and the hairpin; and

the method further comprises detecting the label after the washing.

7. The method of claim 5, further comprising removing the ligated QC probes after the detecting of the label.

8. The method of claim 7, wherein:

the hairpins each comprise a stem comprising complementary first and second single strands and a loop, the first single strand having a first end coupled with the loop, a second end coupled with a first end of the template portion, and a sequence specific to a nuclease, the second single strand having a first end coupled with the loop and a sequence specific to a nuclease; and

removing of the ligated QC probes is performed using a nuclease that binds to the specific sequences.

9. A method of assessing biochip quality, the method comprising:

providing a biochip comprising;

an immobilization layer; and

at least first full length polynucleotide probes and second truncated polynucleotide probes, each having 3′

ends that are coupled to the immobilization layer, the first full length polynucleotide probes having a different number of nucleotide monomers than the second truncated polynucleotide probes;

hybridizing Quality Control (QC) probes to the first full length polynucleotide probe and second truncated polynucleotide probe, each QC probe comprising a template portion having a nucleotide sequence complementary to the entire first full length polynucleotide probe; and

exposing the biochip to conditions suitable for ligation of 3′

ends of the QC probes to 5′

ends of the first full length polynucleotide probe and second truncated polynucleotide probe;

washing the biochip to de-hybridize the QC probes from the first full length polynucleotide probes and second truncated polynucleotide probes, thereby removing un-ligated QC probes;

detecting a ratio of the first amount of the first full length polynucleotide probes that are ligated with the QC probes to the amount of the second truncated polynucleotide probes that are ligated to the QC probes to assess the quality of the biochip, wherein the quality of the biochip is a measure of the relative amounts of the first full length and second truncated polynucleotide probes.

10. The method of claim 9, wherein detecting comprises:

annealing the ligated QC probes and the first full length polynucleotide probes to produce double-stranded structures; and

intercalating a label into the double-stranded structures.

11. The method of claim 9, further comprising completely removing the ligated QC probes from the biochip.