摘要:
The current invention is directed to a method for obtaining a nucleic acid encoding an immunoglobulin variable domain from a single cell comprising the following steps:—performing a first polymerase chain reaction with three to six 5′-primer and one 3′-primer, performing with the product of the first polymerase chain reaction a second polymerase chain reaction with thirteen to sixteen 5′-primer and one 3′-primer, whereby the distance of the binding locations of the primer employed in the second polymerase chain reaction is reduced compared to the first polymerase chain reaction.
摘要:
The current invention is directed to a method for obtaining a nucleic acid encoding an immunoglobulin variable domain from a single cell comprising the following steps: —performing a first polymerase chain reaction with three to six 5′-primer and one 3′-primer, performing with the product of the first polymerase chain reaction a second polymerase chain reaction with thirteen to sixteen 5′-primer and one 3′-primer, whereby the distance of the binding locations of the primer employed in the second polymerase chain reaction is reduced compared to the first polymerase chain reaction.
摘要:
Disclosed is a fluorescent dye comprising a benzothiazolium moiety and a pyrimidinium moiety connected by a mono-methine bridge, characterized in that (i) the 2-position of the pyrimidine carries a substituent which starts with a C-atom and (ii) the 5- and 6- positions of the pyrimidine ring are an integral part of a further aromatic ring structure as exemplified by the following:
摘要:
The present invention provides kits and methods for composition ratio control based on dyes that are designed to enable energy transfer between each other. In more detail, with the method of the present invention it is possible to verify the mixing ratio of a first component comprising a first dye with a second component comprising a second dye.
摘要:
The current invention restates substituted indole nucleosides as both terminal as well as internal building blocks of labeled oligonucleotide probes for the detection, analysis and quantitation of nucleic acids. The substituent comprises a linker and a detectable group or a linker and a reactive group for post synthesis coupling. These modified nucleosides grant access to a wide application area. These new substituted indole nucleosides can be used as labeling reagents for the facile preparation of e.g. optimized hybridization probes, Taqman-probes, or molecular beacon probes.
摘要:
The present invention provides kits and methods for composition ratio control based on dyes that are designed to enable energy transfer between each other. In more detail, with the method of the present invention it is possible to verify the mixing ratio of a first component comprising a first dye with a second component comprising a second dye.
摘要:
The current invention restates substituted indole nucleosides as both terminal as well as internal building blocks of labeled oligonucleotide probes for the detection, analysis and quantitation of nucleic acids. The substituent comprises a linker and a detectable group or a linker and a reactive group for post synthesis coupling. These modified nucleosides grant access to a wide application area. These new substituted indole nucleosides can be used as labeling reagents for the facile preparation of e.g. optimized hybridization probes, Taqman-probes, or molecular beacon probes.
摘要:
The present invention is directed to a fluorescent dye comprising a benzothiazolium moiety and a pyrimidinium moiety connected by a mono-methine bridge, characterized in that (i) the 2-position of the pyrimidine carries a substituent which starts with a C-atom and (ii) the 5- and 6-positions of the pyrimidine ring are an integral part of a further aromatic ring structure.
摘要:
The present invention concerns methods for measuring a nucleic acid amplification in real-time comprising (i) providing a real-time PCR instrument containing a reaction vessel holder for holding multiple reaction vessels and means for positioning the multiple reaction vessels in a reaction vessel holder relative to the detection unit, (ii) filling multiple reaction vessels with a reaction mixture containing thermostable polymerase, deoxynucleotides and buffer, at least two amplification primers, at least one fluorescently-labelled hybridization probe and a fluorescent dye component which is present in a free form during and after the entire amplification reaction, (iii) determining the position of maximum fluorescence emission of the fluorescent dye component in multiple reaction vessels as a function of their position relative to the detection unit, and (iv) performing the amplification reaction and measuring the fluorescence emission of the at least one hybridization probe during and optionally after the amplification reaction for each reaction vessel at the position of maximum fluorescence emission.