Abstract: The application relates to a method of performing a multi-step reaction vessel (68) having at least two compartments (685, 680). The reagents are placed in the first compartment (685) and moved to second one (680) by centrifugation, after which another set of reagents may be placed in the first compartment (685) while the reaction in the lower chamber takes place. Once the reaction is complete, the reagents that were in the first compartment (685) may be moved to the lower one (680) by centrifugation. The application also claims a container having a pierceable lower surface and an upper surface with either a pierceable component or a lid. A wand capable of being electrostatically charged, an apparatus comprising such a wand and a method of transferring solid reagents using such a wand is also claimed.

Abstract: A method for detecting the presence or absence of a polymorphism at a particular position within a nucleic acid sequence in a sample, said method comprising i) annealing a pair of probes to said nucleic acid sequence, said probes being designed such that they anneal adjacent each other, and only completely anneal to one form of the sequence, in a reaction mixture which is substantially free of ATP; ii) ligating together any completely annealed pairs of probes in the presence of NAD and a nucleic acid ligase which uses NAD as a substrate/ iii) phosphorylating any AMP released by a ligation event to ATP, and iv) detecting ATP, in particular using a bioluminescent reaction, in the resultant reaction mixture. Kits for conducting this method are also described and claimed.

Abstract: A method of carrying out an amplification reaction, said method comprising supplying to a well in a disposable unit (a) a sample which contains or is suspected of containing a target nucleic acid sequence (b) primers, nucleotides and enzymes required to effect said amplification reaction and (c) a buffer system, and subjecting the unit to thermal cycling conditions such that any target nucleic acid present within the sample is amplified; wherein the disposable unit comprises a thermally conducting layer and a facing layer having one or more reagent wells of up to 1000 microns in depth defined therebetween; and the reaction mixture comprises at least one of the following: A) a buffer system wherein the pH is above 8.3; B) a detergent; and/or C) a blocking agent. Apparatus for effecting the method as well as disposable units for use in the method are described. The method is particularly suitable for rapid PCR reactions.

Abstract: A method for detecting viable cells such as bacterial cells, within a sample, said method comprising (i) incubating said sample with a virus which is able to infect said cells under conditions which allow said virus to infect and replicate within any such cells which are viable; (ii) detecting any nucleic acid obtained by replication of the virus in said cell.

Abstract: A method for adding a first and a second functional nucleic acid sequence to a reaction mixture, in particular an amplification reaction mixture in a predetermined stoichiometry and/or at a predetermined point in time, said method comprising adding to the reaction mixture an oligonucleotide comprising a first and a second functional nucleic acid sequence separated by a spacer sequence, said spacer sequence comprising a region which, when double stranded, comprises a cleavable region, forming a cleavable double stranded region within the spacer region of said oligonucleotide, and cleaving the double stranded region within said oligonucleotide. Oligonucleotides for use in the method, and comprising a first and a second functional nucleic acid sequence, such as primers or probes used in an amplification reaction, separated by a spacer sequence, is also provided.

Abstract: A method of carrying out an amplification reaction, said method comprising supplying to a well in a disposable unit (a) a sample which contains or is suspected of containing a target nucleic acid sequence (b) primers, nucleotides and enzymes required to effect said amplification reaction and (c) a buffer system, and subjecting the unit to thermal cycling conditions such that any target nucleic acid present within the sample is amplified; wherein the disposable unit comprises a thermally conducting layer and a facing layer having one or more reagent wells of up to 1000 microns in depth defined therebetween; and the reaction mixture comprises at least one of the following: A) a buffer system wherein the pH is above 8.3; B) a detergent; and/or C) a blocking agent. Apparatus for effecting the method as well as disposable units for use in the method are described. The method is particularly suitable for rapid PCR reactions.

Abstract: Method and apparatus for carrying out a thermal cycling reaction, wherein a succession of samples is conveyed through a series of sequentially arranged temperature control sites, each of the sites comprising means for supplying an electric current to, or inducing an electric current in, sample-containing vessels passing through it so as to induce temperature changes in the samples. Also provided is a sample support and its production, the support comprising a succession of sample vessels arranged sequentially one behind the next, preferably in the form of a linked chain, the support comprising an electrically conducting, preferably plastics, material which beats when an electric current passes through it.

Abstract: Analytical methods using RNA probes for the detection or analysis of nucleic acid sequences is described. These probes are contacted with a sample suspected of containing the nucleic acid sequence and if they form duplexes, they are hydrolysed. This may be done, for example during an amplification reaction. AMP generated as a result of the hydrolysis is converted to ATP. The ATP may then be detected using bioluminescent reagents.

Abstract: A method of determining the length of a particular region within a nucleic acid, the method comprising a) subjecting a sample of the nucleic acid to a plurality of amplification reactions in which the region is amplified, wherein the time of the extension phase in each of the reactions is varied; b) monitoring the progress of the amplification reactions; c) determining the minimum time during which extension phase of the amplification is completed within each reaction mixture and relating that to the length of the sequence undergoing extension. The method, combined with melting point analysis, will allow percentage GC content of a sequence to be determined. Length analysis of this type can be used in diagnosis or analysis as well as in recombinant DNA technology to check for the presence of concatamers, and in taxonomic classification or forensics. Apparatus for use in the method is also described and claimed.

Abstract: A method for detecting the presence of a lysed eukaryotic cell in a sample, said method comprising: (i) adding adenosine diphosphate (ADP) to said sample under conditions which allows the conversion of ADP to adenosine triphosphate (ATP) by cellular adenylate kinase, (ii) detecting ATP in said sample and relating that to the presence of adenylate kinase and thus to the presence of lysed cells. The method is useful in detecting the cell lysis, for example when screening for drugs which are required to cause lysis, for example for use in tumor therapy. However, in addition, cells may, in a preliminary step, be lysed and the contents quantitated in order to establish for example the health of condition of the cells or to detect the presence of cells in sample such as milk or urine, for diagnostic purposes.

Abstract: An air sampler for use in analyzing biological or other analytes, comprizes an airflow chamber having an air inlet and an air outlet. An extract from an air mover draws ambient air in through the inlet, and then through the chamber, to discharge it back to atmosphere by way of an outlet. The airflow chamber houses a cyclone air/liquid separator through which the inflowing air is caused to pass. An injector introduces liquid into air flowing through the air inlet. The cyclone separator extracts this liquid and any entrained particles therein and discharges it from the airflow chamber, initially by way of a duct, where it is then conveyed to a collection vessel, by way of a three channel peristaltic pump, and a duct. The collection vessel is housed within a Peltier-cooled holder to maintain the contents of the vessel within the 2-8 degree Celsius range.

Abstract: A method is provided for determining the presence of a target bacteria based on its resistance to a cell lysing antibiotic. Said antibiotic is used to lyse cells of non-target bacteria in a sample and hence facilitate isolation of the target prior to detection by known means.

Abstract: A delivery apparatus for selectively delivering one or more liquid reagents into a reaction or test chamber (2), especially of an assay apparatus, the apparatus comprising: one or more respective storage chambers (5,6) for containing the one or more liquid reagents and arranged generally above the reaction or test chamber (2); and a plunger element (4) arranged and operable for insertion into the mouth of a selected storage chamber so as to displace a selected reagent from therewithin into the reaction or test chamber (2) generally therebelow by gravitational liquid overflow from the mouth of the chamber. The apparatus may conveniently be provided as a discrete delivery unit, with the storage chambers (5,6) prefilled with the selected reagents.

Abstract: Analytical methods using RNA probes for the detection or analysis of nucleic acid sequences is described. These probes are contacted with a sample suspected of containing the nucleic acid sequence and if they form duplexes, they are hydrolysed. This may be done, for example during an amplification reaction. AMP generated as a result of the hydrolysis is converted to ATP. The ATP may then be detected using bioluminescent reagents.

Abstract: The surface plasmon resonance apparatus (1) for detecting a soluble and/or particulate analyte comprises a prism sensor (2) providing a metallised sensor surface (3) capable of binding the analyte, and a laser (10) providing a beam (4) for direction at the sensor surface (3). A detector (5) is provided which is capable of detecting light (6), which is internally reflected from the sensor surface (3). Displacement means (7) comprising a vibrator (8) and mirror (9), is operable whereby the excitation beam (4) is displaced over an angular range (A) relative to the sensor surface (3). An analyte sample (15) is disposed on the metallic sensor (3). The detector (5) is used to interpret the light signals (6) internally reflected from the sensor surface (3) so that the sample (15) is analysed. A video camera system (16), which includes a mirror (17), microscope objective (18) and CCD (charge-coupled device) array (19), is used to detect the scattered light emitted from the upper face of the sensor surface (3).

Abstract: With reference to FIG. 1, and air sampler 1 for use in analysing biological or other analytes, comprises an airflow chamber 2 having an air inlet 3 and an air outlet 4. An extract from 5, draws ambient air in through the inlet 3, and then through the chamber 2, to discharge it back to atmosphere by way of an outlet 6. The airflow chamber 2 houses a cyclone air/liquid separator 7 through which the inflowing air is caused to pass. An injector 8 introduces liquid into air flowing through the air inlet 3. The cyclone separator 7 extracts this liquid and any entrained particles therein and discharges it from the airflow chamber 2, initially by way of a duct 9, where it is then conveyed to a collection vessel 12, by way of a three channel peristaltic pump 13, and a duct 14. The collection vessel 12 is housed within a Peltier-cooled holder 15 to maintain the contents of the vessel 12 within the 2° C.-8° C.

Abstract: Method and apparatus for carrying out a thermal cycling reaction, wherein a succession of samples is conveyed through a series of sequentially arranged temperature control sites, each of the sites comprising means for supplying an electric current to, or inducing an electric current in, sample-containing vessels passing through it so as to induce temperature changes in the samples. Also provided is a sample support and its production, the support comprising a succession of sample vessels arranged sequentially one behind the next, preferably in the form of a linked chain, the support comprising an electrically conducting, preferably plastics, material which beats when an electric current passes through it.

Abstract: A method of analysing the length of a particular region within a nucleic acid, said method comprising a) subjecting a sample of said nucleic acid to a plurality of amplification reactions in which said region is amplified, wherein the time of the extension phase in each of said reactions is varied; b) monitoring the progress of said amplification reactions; c) determining the minimum time during which extension phase of the amplification is completed within each reaction mixture and relating that to the length of the sequence undergoing extension. The method, combined with melting point analysis will allow percentage GC content of a sequence to be determined. Length analysis of this type can be used in diagnosis or analysis as well as in recombinant DNA technology to check for the presence of concatamers, and in taxonomic classification or forensics. Apparatus for use in the method is also described and claimed.

Abstract: The surface plasmon resonance apparatus (1) for detecting a soluble and/or particulate analyte comprises a prism sensor (2) providing a metallized sensor surface (3) capable of binding the analyte, and a laser (10) providing a beam (4) for direction at the sensor surface (3) A detector (5) is provided which is capable of detecting light (6), which is internally reflected from the sensor surface (3). Displacement means (7) comprising a vibrator (8) and mirror (9), is operable whereby the excitation beam (4) is displaced over an angular range (A) relative to the sensor surface (3). An analyte sample (15) is disposed on the metallic sensor (3). The detector (5) is used to interpret the light signals (6) internally reflected from the sensor surface (3) so that the sample (15) is analyzed. A video camera system (16), which includes a mirror (17), microscope objective (18) and CCD (charge-coupled device) array (19), is used to detect the scattered light emitted from the upper face of the sensor surface (3).

Abstract: Enzymes and methods suitable for assaying ATP, and specific application for such assays are described and claimed. In particular, there is described a recombinant mutant luciferase having a mutation for example, in the amino-acid corresponding to amino acid residue number 245 in Photinus pyralis, is such that the Km for ATP of the luciferase is increased e.g. five-fold with respect to that of the corresponding non-mutated enzyme such that it is of the order of 500 &mgr;m-1 mM. Also disclosed are luciferases having additional mutations conferring improved thermostability or altered wavelength of emitted light. Recombinant polynucleotides, vectors and host cells are also disclosed, as are methods of assaying the amount of ATP in a material (e.g. cells) optionally in real-time. Also disclosed are test-kits for in vitro assays.