Chemistry: molecular biology and microbiology – Apparatus – Including condition or time responsive control means
Patent
1993-07-22
1996-01-02
Jones, W. Gary
Chemistry: molecular biology and microbiology
Apparatus
Including condition or time responsive control means
204299R, 435 4, 435 6, 4352865, 4352872, C25D 112
Patent
active
054808030
DESCRIPTION:
BRIEF SUMMARY
BACKGROUND OF THE INVENTION
This application is the national filing of PCT application No. PCT/GB91/00308.
It is common practice in the fields of biomedicine and molecular biology to use techniques for the analysis of biological molecules where the analytes are immobilised on the surface of membrane filters. Such techniques are well documented in the scientific literature and have been used for the analysis of DNA, RNA and protein molecules. The processes used in such analyses are commonly termed Southern blotting (1) where DNA molecules or fragments are analysed, Northern blotting (2) where RNA molecules or fragments are analysed and Western blotting (3) where proteins or polypeptides are analysed.
The principle of all these techniques involves separation of the analyte molecules by electrophoresis in agarose or acrylamide gels such that the molecules or fragments of molecules are separated according to their molecular weight or physical size. Separation is followed by transfer of the analyte to a membrane composed of natural or synthetic polymer to produce an exact replica of the separation pattern on the gel. Such transfer is commonly achieved by capillary transfer, application of an electric field or by use of vacuum suction.
In addition to the above techniques there are a variety of further blotting procedures in common use in biomedicine and molecular biology. The technique commonly known as the unblot (4) process involves drying the gel used for separation of the analyte to form a thin sheet which can then be handled and treated identically to a membrane blot. The techniques known as dot blotting (5) and slot blotting (6) are also widely used to confirm the presence or absence of a particular molecule or molecular fragment in an unseparated analyte mixture. In these techniques the analyte is applied directly to the membrane filter, either by use of a pipette tip, or by using a manifold filtration device.
Further blotting techniques are used in the process of gene cloning, to identify organisms containing an inserted nucleic acid sequence. In these procedures, known as colony lifts where bacteria are the organism used, or plaque lifts where the organism is viral, a filter is laid on the surface of a culture plate containing the organism and then removed, forming a replica of the distribution of organisms on the culture plate, which is then analysed for the presence or absence of the inserted nucleic acid sequence.
Central to all of the above techniques is the requirement to expose the immobilised analyte to a solution containing a further biomolecule, commonly termed a probe, which has an affinity for one or more analyte molecules, such that the probe molecule is able to bind to the immobilised analyte.
The probe molecule, which may be DNA, RNA, a synthetic oligonucleotide, a protein or polypeptide, or a ligand, is labelled with a radioactive isotope or non-radioactive reporter molecule, which may be either an enzyme or a small molecule (typically biotin or a hapten) recognised by an antibody or other binding protein.
Following exposure to the probe the blot is washed in order to achieve removal of unbound probe. Where the probe is labelled with a radioisotope, exposure of the blot to X-ray film produces an image on the film corresponding to the location of the analyte molecule or molecules recognised by the probe.
Where the probe carries a non-isotopic label further stages are necessary to achieve localisation of the bound probe. In the case of enzyme labelled probes, addition of a specialised enzyme substrate solution leads to the formation of a coloured stain or the emission of light at the position of the probe. Where the probe is labelled with biotin or a hapten molecule the blot is exposed to a solution containing either avidin or steptavidin in the case of biotin labelled probes, or to a solution containing an antibody recognising the hapten molecule. The antibodies or biotin binding proteins used in these procedures may themselves be labelled with either a radioisotope or an enzyme label, an
REFERENCES:
patent: 2031010 (1936-02-01), Simjian
patent: 3435835 (1969-04-01), Hobbs
patent: 4222843 (1980-09-01), Suzuki et al.
patent: 4248514 (1979-10-01), Watkins
patent: 4982326 (1991-01-01), Kaneko
Amersham International plc
Houtteman Scott
Jones W. Gary
LandOfFree
Apparatus for treating analytes does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Apparatus for treating analytes, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Apparatus for treating analytes will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-234595