Process for improving efficiency of DNA amplification reaction

• US 20040110182A1
• Filed: 06/20/2003
• Published: 06/10/2004
• Est. Priority Date: 01/10/2002
• Status: Abandoned Application

• Current Assignee: Nichirei Corporation
• Original Assignee: Nichirei Corporation
• Inventors: Yamamoto, Satoshi, Koizumi, Takeshi, Hamano, Yoko
• Application Number:
  US10/601,713
• Publication Number:
  US 20040110182A1
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: C12Q 1/6832 Enhancement of hybridisation reaction
• CPC Class Codes: C12Q 1/6853 using modified primers or templates
• CPC Class Codes: C12Q 1/686 Polymerase chain reaction [PCR]
• CPC Class Codes: C12Q 2525/161 incorporating target specific and non-target specific sites
• CPC Class Codes: C12Q 2531/107 Probe or oligonucleotide ligation
• CPC Class Codes: C12Q 2563/107 fluorescence
• CPC Class Codes: C12Q 2563/131 the label being a member of a cognate binding pair, i.e. extends to antibodies, haptens, avidin

Abstract

The present invention has an object of providing a process for improving the efficiency of a DNA amplification reaction, and a process for improving the hybridization specificity of an oligonucleotide to a DNA.

The present invention provides a process for improving the efficiency of a DNA amplification reaction, wherein a primer in which a compound such as LC-Red 705 or an oligonucleotide with a GC content of at least 25% and with at least four bases is added to the 5′ terminus is used as the primer; as well as a process for improving the hybridization specificity of an oligonucleotide to a DNA sample, wherein an oligonucleotide in which a compound such as LC-Red 705 is conjugated to the 5′ terminus is used for hybridizing to a DNA.

Claims

1. A process for improving efficiency of a DNA amplification reaction, wherein a primer, in which a compound selected from a group consisting of LC-Red 705, an amino group, a phosphate group, biotin, DIG, DNP, TAMRA, Texas-Red, ROX, XRITC, rhodamine, LC-Red 640, a mercapto group, psoralen, cholesterol, FITC, 6-FAM, TET, cy3, cy5, BODIPY 564/570, BODIPY 500/510, BODIPY 530/550, BODIPY 581/591 and oligonucleotides with a combined G and C content of at least 25% and with at least four bases is added to a 5′

terminus, is used as a primer.

2. A process for improving efficiency of a DNA amplification reaction according to claim 1, wherein said oligonucleotide with a combined G and C content of at least 25% and with at least four bases has a combined G and C content of at least 50%, comprises no more than 40 bases, and has a quantity of a more numerous base of G and C that accounts for at least 50% of said combined G and C content, and a quantity of a more numerous base of A and T that accounts for at least 50% of a combined content of A and T.

3. A process for improving efficiency of a DNA amplification reaction according to either one of claim 1 and claim 2, which is a process for improving PCR amplification efficiency.

4. A process for improving efficiency of a DNA amplification reaction according to claim 3, wherein said PCR is either one of asymmetric PCR and degenerate PCR.

5. A process for improving hybridization specificity of an oligonucleotide to a DNA, wherein an oligonucleotide in which a compound selected from a group consisting of LC-Red 705, an amino group, a phosphate group, biotin, DIG, DNP, TAMRA, Texas-Red, ROX, XRITC, rhodamine, LC-Red 640, a mercapto group, psoralen, cholesterol, FITC, 6-FAM, TET, cy3, cy5, BODIPY 564/570, BODIPY 500/510, BODIPY 530/550 and BODIPY 581/591 is conjugated to a 5′

terminus is used for hybridizing to said DNA.