Patent classifications
C12Q2521/319
Nucleic acid enrichment and detection
Disclosed is a hybridisation capture method based on the pyrophosphorolysis reaction. According to the present invention, there is provided a method for increasing the ratio of a first nucleic add sequence to second nucleic add sequence in a sample.
METHODS FOR CIRCULARIZING LINEAR DOUBLE STRANDED NUCLEIC ACIDS
A method, in particular an in vitro method, for the circularization of a double stranded DNA nucleic acid. Also, a circularized double stranded DNA nucleic acid obtainable by the method. Another aspect pertains to a host cell comprising a circularized double stranded DNA nucleic acid obtainable by the method. Further, therapeutic and non-therapeutic uses of a circularized double stranded DNA nucleic acid obtainable by the method. Finally, a kit for the circularization of a double stranded DNA nucleic acid.
METHODS FOR CIRCULARIZING LINEAR DOUBLE STRANDED NUCLEIC ACIDS
A method, in particular an in vitro method, for the circularization of a double stranded DNA nucleic acid. Also, a circularized double stranded DNA nucleic acid obtainable by the method. Another aspect pertains to a host cell comprising a circularized double stranded DNA nucleic acid obtainable by the method. Further, therapeutic and non-therapeutic uses of a circularized double stranded DNA nucleic acid obtainable by the method. Finally, a kit for the circularization of a double stranded DNA nucleic acid.
Template-Free Enzymatic Polynucleotide Synthesis Using Photocleavable Linkages
The present invention is directed to methods and kits for template-free enzymatic synthesis of polynucleotides employing photocleavable linkages. In some embodiments, such methods include using 3′-O—NH2-dNTP monomers which may react with photocleavage products having free ketone to allow synthesis and purification on the same or an added support.
Template-Free Enzymatic Polynucleotide Synthesis Using Photocleavable Linkages
The present invention is directed to methods and kits for template-free enzymatic synthesis of polynucleotides employing photocleavable linkages. In some embodiments, such methods include using 3′-O—NH2-dNTP monomers which may react with photocleavage products having free ketone to allow synthesis and purification on the same or an added support.
METHOD FOR EVALUATING VIRAL CLEARANCE CAPABILITY
Provided is a method for evaluating the viral clearance capability of a virus removal medium, comprising the steps of: (a) adding a viral capsid-containing liquid to a solution to be purified; (b) contacting the virus removal medium with the solution to be purified that has been supplemented with the viral capsid-containing liquid to harvest a purified solution; and (c) quantifying a total viral capsid in the solution to be purified before the purification and a total viral capsid in the purified solution.
METHOD FOR EVALUATING VIRAL CLEARANCE CAPABILITY
Provided is a method for evaluating the viral clearance capability of a virus removal medium, comprising the steps of: (a) adding a viral capsid-containing liquid to a solution to be purified; (b) contacting the virus removal medium with the solution to be purified that has been supplemented with the viral capsid-containing liquid to harvest a purified solution; and (c) quantifying a total viral capsid in the solution to be purified before the purification and a total viral capsid in the purified solution.
MICROARRAY SYSTEM WITH IMPROVED SEQUENCE SPECIFICITY
The invention provides a novel array method for nucleic acid sequence detection with improved specificity which allows for detection of genetic variation, from simple SNPs (where the variation occurs at a fixed position and is of limited allelic number) to more complex sequence variation patterns (such as with multigene families or multiple genetic strains of an organism where the sequence variation between the individual members is neither fixed nor consistent). The array is comprised of short, synthetic oligonucleotide probes attached to a solid surface which are hybridized to single-stranded targets. Single stranded targets can be produced using a method that employs primers modified on the 5′ end to prohibit degradation by a 5′-exonuclease that is introduced to degrade the unprotected strand. The invention further provides for printing buffers/solutions for the immobilization of oligonucleotide probes to an array surface. The invention also provides hybridization and wash buffers and conditions to maximize hybridization specificity and signal intensity, and reduce hybridization times.
MICROARRAY SYSTEM WITH IMPROVED SEQUENCE SPECIFICITY
The invention provides a novel array method for nucleic acid sequence detection with improved specificity which allows for detection of genetic variation, from simple SNPs (where the variation occurs at a fixed position and is of limited allelic number) to more complex sequence variation patterns (such as with multigene families or multiple genetic strains of an organism where the sequence variation between the individual members is neither fixed nor consistent). The array is comprised of short, synthetic oligonucleotide probes attached to a solid surface which are hybridized to single-stranded targets. Single stranded targets can be produced using a method that employs primers modified on the 5′ end to prohibit degradation by a 5′-exonuclease that is introduced to degrade the unprotected strand. The invention further provides for printing buffers/solutions for the immobilization of oligonucleotide probes to an array surface. The invention also provides hybridization and wash buffers and conditions to maximize hybridization specificity and signal intensity, and reduce hybridization times.
Detection of target nucleic acid sequences by PO cleavage and hybridization
The present invention relates to the detection of a target nucleic acid sequence by a POCH (PO Cleavage and Hybridization) assay on a solid substrate. The present invention detects the target nucleic acid sequence by use of in which the PO (Probing Oligonucleotide) hybridized with the target nucleic acid sequence is cleaved and the cleavage of the PO is detected by hybridization with the CO (Capturing Oligonucleotide). In the present invention, an uncleaved PO is hybridized with the CO immobilized onto the solid substrate. The designs of the PO and the CO are convenient and the optimization of reaction conditions is routinely easy in the present invention. Where the detection of signal on the solid substrate is continuously performed along with repetition of cleavage of the POs in the present invention, the number of the POs cleaved is increased upon the repetition number of the cleavage reaction and the signal is changed in parallel with the number of the POs cleaved. Then, the target nucleic acid sequence can be detected in a real-time manner. In contrast, the change of the signal is not observed in the absence of the target nucleic acid sequence.