Abstract: The present invention relates to the detection of a target nucleic acid sequence by a PTOCE (PTO Cleavage and Extension) assay. The present invention detects a target nucleic acid sequence in which the PTO (Probing and Tagging Oligonucleotide) hybridized with the target nucleic acid sequence is cleaved to release a fragment and the fragment is hybridized with the CTO (Capturing and Templating Oligonucleotide) to form an extended duplex, followed by detecting the presence of the extended duplex. The extended duplex provides signals (generation, increase, extinguishment or decrease of signals) from labels indicating the presence of the extended duplex and has adjustable Tm value, which are well adoptable for detection of the presence of the target nucleic acid sequence.

Abstract: The present invention relates to detection of a target nucleic acid sequence in a sample using two different detection temperatures. The present invention using difference between signals detected at two detection temperatures enables to decrease well-to-well variation or sample-to-sample variation generated in real-time PCR processes in more convenient and effective manner.

Abstract: The present invention relates to the detection of a target nucleic acid sequence by a PTOCE (PTO Cleavage and Extension) assay. The present invention detects a target nucleic acid sequence in which the PTO (Probing and Tagging Oligonucleotide) hybridized with the target nucleic acid sequence is cleaved to release a fragment and the fragment is hybridized with the CTO (Capturing and Templating Oligonucleotide) to form an extended duplex, followed by detecting the presence of the extended duplex. The extended duplex provides signals (generation, increase, extinguishment or decrease of signals) from labels indicating the presence of the extended duplex and has adjustable Tm value, which are well adoptable for detection of the presence of the target nucleic acid sequence.

Abstract: The present invention relates to a method for quantifying a target nucleic acid sequence by use of a common internal control. The present invention allows to determine an absolute initial amount of a target nucleic acid sequence with no use of a standard curve. The present invention amplifies not only a control reaction mixture comprising a known-amount standard and an internal control but also a sample reaction mixture comprising an internal control identical to the internal control in the control reaction mixture and the target nucleic acid sequence to be quantified. In the present invention, a relative ratio of the amount of the standard and the amount of the target nucleic acid sequence is calculated by using the internal control and then an initial amount of the target nucleic acid sequence is determined by using a known amount of the standard.

Abstract: The present invention relates to a novel method for detection of target nucleic acid sequences by cyclic exonucleolytic reactions (CER) or exonucleolytic reactions (ER) using single-labeled immobilized probes on a solid phase. The present invention enables to detect target nucleic acid sequences on a solid phase using single-labeled systems. Comparing with multiple-labeled systems such as dual labeling, the present invention using single-labeled probes has excellent advantages in light of convenience and cost effectiveness in probe design and preparation. Furthermore, the measurement of changes of the signal decrease during reactions is responsible for more accurate qualitative and quantitative analysis of target nucleic acid sequences.

Abstract: The present invention relates to the detection of a target nucleic acid sequence by a PTOCE (PTO Cleavage and Extension) assay. The present invention detects a target nucleic acid sequence in which the PTO (Probing and Tagging Oligonucleotide) hybridized with the target nucleic acid sequence is cleaved to release a fragment and the fragment is hybridized with the CTO (Capturing and Templating Oligonucleotide) to form an extended duplex, followed by detecting the presence of the extended duplex. The extended duplex provides signals (generation, increase, extinguishment or decrease of signals) from labels indicating the presence of the extended duplex and has adjustable Tm value, which are well adoptable for detection of the presence of the target nucleic acid sequence.

Abstract: The present invention is generally drawn to a novel method for determining a SNP (single nucleotide polymorphism) genotype using a PTO-SNV (Probing and Tagging Oligonucleotide for Single Nucleotide Variation). The present invention provides novel protocols for SNP genotyping in which only one allele-specific oligonucleotide permits in a SNP genotyping reaction to determine whether a target nucleic acid sequence to be analyzed is homozygous or heterozygous for the SNP allele of interest or has no SNP allele of interest.

Abstract: The present invention relates to the detection of a target nucleic acid sequence by a cyclic exonucleolytic reaction. The present method enabling to generate signals by probe digestion with no help of primers and to amplify signals with no help of simultaneous target amplification reactions may enable to detect multiple target sequences without any problems accounted in the conventional real-time PCR methods such as false positive signals and difficulties in oligonucleotides (primer and probe) selection and reaction condition optimization.

Abstract: The present invention relates to the detection of a target nucleic acid sequence from a DNA or a mixture of nucleic acids by a PCE-hCTO (PTO Cleavage and Extension using hCTO) assay on a solid phase. According to the present invention, the extended duplex is formed in a liquid phase in a target-dependent manner and then its presence is detected on a solid phase. Since hCTO is not immobilized onto a solid phase, the extended duplex is more effectively formed in a liquid phase.

Abstract: The present invention relates to the detection of a target nucleic acid sequence by a PCE-IH (PTO Cleavage and Extension-Dependent Immobilized Oligonucleotide Hybridization) assay on a solid phase. The present invention firstly hybridizes the PTO with a target nucleic acid sequence, forms the extended strand in a target-dependent manner by using the CTO having artificially selected sequence as templates and finally hybridizes the extended strand with the IO immobilized on a solid phase. In other words, the present invention employs a series of reactions including PTO hybridization and cleavage, CTO hybridization and extension and IO hybridization, which is responsible for the highly enhanced specificity of the present invention.

Abstract: The present invention relates to the detection of a nucleotide variation on a target nucleic acid sequence using an amplification blocker and a VD-PTOCE (Variation Detection by PTO Cleavage and Extension) assay. The present invention is significantly effective in the detection of a minority mutation in an excess of wild-type DNA.

Abstract: The present invention relates to the detection of a target nucleic acid sequence using a target hybridization and detection primer (THD primer). The present invention allows for both a target amplification and a signal amplification by introducing a label into a primer used in PCR reactions, ensuring a real-time target detection by PCR reaction by no use of complicated oligonucleotides. The present invention could completely be free from the troublesome matters and shortcomings associated with conventional real-time PCR methods. The present invention allows for successful real-time target detection by using only a labeled primer. This feature makes it possible that the present invention exhibits excellent real-time target detection in multiplex manner.

Abstract: The present invention relates to a method for quantifying a target nucleic acid sequence performed in such a manner that at least two cycles in the nucleic acid amplification subject to melting peak analysis are predetermined before the nucleic acid amplification and melting peak analyses are performed for the at least two predetermined cycles, followed by quantifying the target nucleic acid sequence using data values from the melting peak curve (e.g., the presence or absence, height and area).

Abstract: The present invention relates to the detection of a target nucleic acid sequence by a PCE-NH (PTO Cleavage and Extension-Dependent Non-Hybridization) assay. The present invention adopts the occurrence of the inhibition of the hybridization between the HO with the CTO by the formation of the target-dependent extended duplex. Therefore, the present invention may detect target sequences even when the HO is not cleaved. In this regard, the design of the 5?-tagging portion of PTO, CTO and HO sequences may be readily performed and the conditions for reactions may be also easily established. In addition, the detection of the hybrid between the CTO and the HO may be performed in a different vessel from that for the extension of the CTO.

Abstract: The present invention relates to the detection of a target nucleic acid sequence by a PTO Cleavage and Extension-Dependent Signaling Oligonucleotide Cleavage assay (PCE-SC assay). The present invention is carried out in such a manner that the extended strand is produced on the CTO having arbitrary sequences as templates depending on the presence of target nucleic acid sequences and in turn the SO as probes is hybridized with the extended strand to give signal. The present invention employs a series of reactions including PTO hybridization and cleavage, CTO hybridization and extension, and SO hybridization and cleavage, which is responsible for the highly enhanced specificity of the present invention.

Abstract: The present invention is generally drawn to a novel method and a kit for detecting nucleotide variations by a PTOCE (PTO Cleavage and Extension) assay with PTO-NV. Furthermore, the present invention is directed to a novel method and a kit for detecting a nucleotide variation on a target nucleic acid sequence by a PTOCE assay with PTO-NV having a non-base paring moiety.

Abstract: The present invention relates to the detection of a target nucleic acid sequence by a PTOCE (PTO Cleavage and Extension) assay. The present invention detects a target nucleic acid sequence in which the PTO (Probing and Tagging Oligonucleotide) hybridized with the target nucleic acid sequence is cleaved to release a fragment and the fragment is hybridized with the CTO (Capturing and Templating Oligonucleotide) to form an extended duplex, followed by detecting the presence of the extended duplex. The extended duplex provides signals (generation, increase, extinguishment or decrease of signals) from labels indicating the presence of the extended duplex and has adjustable Tm value, which are well adoptable for detection of the presence of the target nucleic acid sequence.

Abstract: The present invention relates to the detection of a target nucleic acid sequence by a PCE-SH (PTO Cleavage and Extension-Dependent Signaling Oligonucleotide Hybridization) assay. The present invention does not use probes to be hybridized with target nucleic acid sequences for providing target signals. Interestingly, the present invention uses probes (signaling oligonucleotides) to be hybridized with the extended strand formed in a target-dependent manner in which the extended strand is synthesized using the CTO artificially selected as templates.

Abstract: The present invention relates to the detection of a target nucleic acid sequence by a PTOCE (PTO Cleavage and Extension) assay. The present invention detects a target nucleic acid sequence in which the PTO (Probing and Tagging Oligonucleotide) hybridized with the target nucleic acid sequence is cleaved to release a fragment and the fragment is hybridized with the CTO (Capturing and Templating Oligonucleotide) to form an extended duplex, followed by detecting the presence of the extended duplex. The extended duplex provides signals (generation, increase, extinguishment or decrease of signals) from labels indicating the presence of the extended duplex and has adjustable Tm value, which are well adoptable for detection of the presence of the target nucleic acid sequence.

Abstract: The present invention relates to an annealing control primer for improving annealing specificity in nucleic acid amplification and its applications to all fields of nucleic acid amplification-involved technology. The present primer comprises (a) a 3?-end portion having a hybridizing nucleotide sequence substantially complementary to a site on a template nucleic acid to hybridize therewith; (b) a 5?-end portion having a pre-selected arbitrary nucleotide sequence; and (c) a regulator portion positioned between said 3?-end portion and said 5?-end portion comprising at least one universal base or non-discriminatory base analog, whereby said regulator portion is capable of regulating an annealing portion of said primer in association with annealing temperature.