90 results about "Displacement activity" patented technology

Methods of amplifying a target nucleic acid whereby the target nucleic acid is amplified in the presence of a deoxyribonucleotide triphosphate, a DNA polymerase having strand displacement activity, at least one chimeric oligonucleotide primer, at least one upstream block oligonucleotide and a RNase H; wherein the chimeric oligonucleotide primer contains a ribonucleotide positioned at the 3′-terminus; wherein the upstream block oligonucleotide is capable of annealing to a region 3′ to a portion in the nucleic acid as the template to which the chimeric oligonucleotide primer anneals; and compositions and kits thereof.

An object to be achieved by the present invention is to provide a nucleic acid amplification method by which a nucleic acid can be amplified substantially isothermally using oligonucleotide primers and DNA polymerase capable of strand displacement. The present invention provides a nucleic acid amplification method which comprises performing substantially isothermal incubation of a reaction solution containing at least one type of deoxynucleotide triphosphate, at least one type of DNA polymerase having strand displacement activity, a divalent cation, at least 0.01% or more surfactant, at least two types of oligonucleotide primer, and the nucleic acid fragment as a template so as to perform a polymerase reaction that initiates from the 3′ end of the primer and thus amplifying the nucleic acid fragment.

Thermostable viral polymerases exhibiting a combination of activities selected from, proofreading (3′-5′) exonuclease activity, nick translating (5′-3′) nuclease activity, synthetic primer-initiated polymerase activity, nick-initiated polymerase activity, reverse transcriptase activity, strand displacement activity, and / or decreased discrimination against incorporation of nucleotide analogs. Also provided are compositions including the polymerases, polynucleotides encoding the polymerases and methods of using the polymerases.

An object to be achieved by the present invention is to provide a nucleic acid amplification method by which a nucleic acid can be amplified substantially isothermally using oligonucleotide primers and DNA polymerase capable of strand displacement. The present invention provides a nucleic acid amplification method which comprises performing substantially isothermal incubation of a reaction solution containing at least one type of deoxynucleotide triphosphate, at least one type of DNA polymerase having strand displacement activity, a divalent cation, at least 0.01% or more surfactant, at least two types of oligonucleotide primer, and the nucleic acid fragment as a template so as to perform a polymerase reaction that initiates from the 3′ end of the primer and thus amplifying the nucleic acid fragment.

The present invention provides a nucleic acid amplification method by using a oligonucleotide primer and a DNA polyase. The nucleic acid amplification method provided by the invention comprises performing incubation of a reaction solution containing at least one type of deoxynucleotide triphosphate, at least one type of DNA polymerase having strand displacement activity, at least two types of oligonucleotide primer, and the nucleic acid fragment as a template so as to perform a polymerase reaction that initiates from the 3' end of the primer and thus amplifying the nucleic acid fragment, wherein the characteristic is that a first oligonucleotide primer and a second oligonucleotide primer are designed in such a way that a region which contains two identical sequences X of serial 4 or more nucleotides within the region of 200 or less nucleotides, or a part thereof can be amplified.

The invention discloses an RT-LAMP (reverse transcription and loop-mediated isothermal amplification) detection primer group, an RT-LAMP detection kit and an RT-LAMP detection method for a simian immunodeficiency virus. The detection primer group comprises a pair of outer primers, a pair of loop primers and a pair of inner primers. The detection kit comprises the primer group, RT-LAMP reaction liquid, Bst DNA polymerase, reverse transcriptase as well as negative control and positive control. The detection method comprises the following steps: extracting to-be-detected virus RNA; conducting reverse transcription on the RNA under the action of revertase; then, amplifying a sample template by virtue of six specific primers and one Bst DNA polymerase having strand displacement activity at 63-65 DEG C; and judging whether a to-be-detected sample contains the simian immunodeficiency virus (SIV) RNA by observing whether a reaction tube solution becomes muddy or not with naked eyes or performing electrophoresis to analyze whether a ladder exists or not. The detection primer group, the detection kit and the detection method provided by the invention have the advantages of being highly specific, high in sensitivity, rapid and efficient, simple and convenient to operate, easy in result reading and the like, and are suitable for popularization and application in grassroots animal epidemic disease monitoring units and experimental monkey breeding enterprises in remote areas.

An object of the present invention is to provide an inexpensive and simple method for synthesis of a double-stranded DNA corresponding to a particular RNA, and a method for amplification of the aforementioned double-stranded DNA. The present invention relates to a method for synthesis of a double-stranded DNA having a nucleotide sequence corresponding to template RNA having polyA, comprising step 1 in which reverse transcription reaction of template RNA is carried out employing oligo(dT)primer to which DNA fragment having a known sequence has been added at the 5'-terminal, to obtain a single-stranded DNA, and step 2 in which double strand formation reaction of single-stranded DNA obtained in step 1 is carried out employing a random primer to which DNA fragment having a known sequence has been added at the 5'-terminal, in the presence of polymerase which does not have 3'->5' exonuclease activity nor strand displacement activity, to obtain a double-stranded DNA, as well as a method for amplification of a double-stranded DNA having a nucleotide sequence corresponding to template RNA having polyA, further comprising step 3 in which PCR reaction is carried out using the obtained double-stranded DNA.

The invention discloses a method for rapidly detecting cow and sheep derived components. The principle is that a cow-derived forward outer primer COW-F3, a cow-derived backward outer primer COW-B3, a cow-derived forward inner primer COW-FIP, a cow-derived backward inner primer COW-FIP, a sheep-derived forward outer primer SHEEP-F3, a sheep-derived backward outer primer SHEEP-B3, a sheep-derived forward inner primer SHEEP-FIP, a sheep-derived backward inner primer SHEEP-BIP and DNA polymerase with the chain displacement activity are used for amplifying nucleic acid at 63-65 DEG C with the amplification efficiency up to 109-1,010 copies in a short time. The identifying method is to directly observe the turbidity of precipitation in a reaction tube by using naked eye or judge that amplification is carried out or not by the color change of added SYBR Green. The detection method has the characteristics of high specificity, high efficiency, rapidness, simple operation, easy detection and the like.

The technology provided herein relates to novel variants of DNA-Polymerases exhibiting high termo-stability as well as a strong strand displacement activity; to nucleic acid molecules encoding said DNA-Polymerases, vectors, host cells containing the nucleic acids and methods for preparation and producing such enzymes; compositions comprising at least one of the DNA-Polymerases; and methods for using such enzymes in DNA sequencing and / or DNA amplification processes.