Chemical apparatus and process disinfecting – deodorizing – preser – Control element responsive to a sensed operating condition
Reexamination Certificate
2000-03-28
2003-07-15
Warden, Jill (Department: 1743)
Chemical apparatus and process disinfecting, deodorizing, preser
Control element responsive to a sensed operating condition
C436S174000, C436S177000, C436S178000, C422S091000, C210S224000, C210S231000, C210S294000, C210S295000
Reexamination Certificate
active
06592826
ABSTRACT:
The present invention relates to a vacuum chamber and a vacuum system using the vacuum chamber for the directed transport of a substance, especially a fluid, and to its use in an apparatus for automatic plasmid preparation.
The past few years have seen an increase in the scale of efforts to obtain the complete genetic information of entire organisms. Beginning with the sequencing of a phage genome (bacteriophage T7: 38000 base pairs, bacteriophage &lgr;: 48514 base pairs) and continuing by way of the genome of
Escherichia coil
(4.2×10
6
base pairs) to the yeast
Saccharomyces cerevisiae
(2.3×10
7
base pairs) as the first representative of the eukaryotes, the number of base pairs to be sequenced has increased almost 600-fold. In the meantime, the human genotype with more than 3×10
9
base pairs has become the goal of these efforts in the “Human Genome Project”. The enormous quantities of DNA to be sequenced are barely manageable by the means and personnel available to laboratories hitherto. There is therefore a demand for new technologies that, for an acceptable financial outlay, are capable of bringing about a considerable increase in the throughput of samples in this research programme. Two mutually influencing strategies have come to light in the course of current development: on the one hand the miniaturisation of laboratory sequences and on the other hand the unsupervised automation of well-established laboratory procedures.
The miniaturisation of laboratory sequences has given rise to miniaturised electrophoresis analysers in which the separation of biomolecules on the basis of their charge and size is utilised. Such miniaturised electrophoresis analysers are obtained by means of microstructures in electrophoresis chips. Also available are miniaturised PCR machines, wherein during the polymerase chain reaction (polymerase chain reaction=PCR) DNA fragments up to 6 kilobases in size are amplified. Also known are miniaturised sample arrays and miniaturised detection systems. Miniaturised elements such as those described above can be combined to form larger units, so that a complete miniaturised laboratory unit is obtained.
On the other hand, for the automation of a laboratory it is not absolutely necessary to miniaturise routine procedures. It is likewise possible to design a robot system that completely or partially replaces the manual tasks of a human being in order to achieve an increase in sample throughput. The following manual tasks are typical of a laboratory preparation (with particular emphasis on plasmid preparation as a preliminary to PCR sequencing) and need to be carried out by suitable robots:
pipetting
transport of used material and of chemicals
suction of fluids through filters, membranes, permeable solids or the like
PCR reaction.
Current pipetting robots locate standard laboratory material on a work surface at defined positions and thus enable tested laboratory protocols to be set up. For example, using auxiliary robots it is possible for microtitre plates, pipette tips or reservoirs for buffer solutions etc. to be installed on such machines and, after use, removed from the workstation again. Thus, all the necessary pipetting steps preliminary to a PCR or a plasmid preparation can be executed in order that the product of that pipetting operation can then be introduced into a suitable machine for further preparation using a gripping robot.
The polymerase chain reaction (PCR) amplifies a DNA segment when it is enclosed between two defined primer sites. If equal amounts of primers are used, double-stranded DNA copies are produced by the PCR, whereas if one primer is used in excess then, in accordance with that excess, single-stranded copies of the amplified DNA are obtained. Both single-stranded and double-stranded DNA can be used for sequencing. In sequencing-intensive projects the DNA fragments to be analysed are cloned into plasmids which are then in the first instance present in a defined matrix of bacterial colonies (Escherichia Coli Blue) growing on agar. The subsequent taking up of the colonies from the matrix into culture tubes can also be automated. Over an incubation period (37° C.) lasting about 12 hours the living bacterial clones then yield sufficient material to obtain in a preparation the plasmid copies necessary for sequencing. Obtaining such purified plasmids for sequencing is achieved, for example, by the QIAWELL 96 ultraplasmid purification procedure. Such plasmid preparation procedures include filtering operations in which a fluid has to be transported in a directed manner from one filter into at least one second filter and either also passes through that filter or is simply collected in a controlled manner.
The problem underlying the invention is therefore to provide an apparatus in which the directed movement of substances can be carried out automatically.
The present invention relates to a vacuum chamber for the directed transport of a substance, especially a fluid, there being installed in the vacuum chamber a permeable means and a collecting means, so that there are defined at least two vacuum regions that can be established independently of one another, namely a first vacuum region between the permeable means and the collecting means and a second vacuum region between the collecting means and the base of the vacuum chamber, and a vacuum can be generated in the two vacuum regions independently of one another so that the substance, especially the fluid, can be sucked from the first permeable means into the collecting means. Fluids are here to be understood as being gases, liquids, vapours and fumes.
The second means is preferably also permeable, so that by the application of a vacuum to the second region the fluid or liquid is sucked through the second means into the second region. The vacuum chamber according to the invention will generally have exactly two vacuum regions, but more than two vacuum regions are possible, for example when several filtrations are to be carried out one immediately after another.
Furthermore, the permeable means are formed by filter supports having a large number of filter elements, so that fluid can be transported in a defined manner from a particular filter element of the first filter support into a corresponding filter element of the second filter support in turn through the latter into the second lower region of the vacuum chamber. The collecting means are likewise collector supports having a defined number of collecting elements. In the case of a collector support, therefore, the substance, especially the fluid, is transported through the first filter support into the collector support.
Moreover, the vacuum chamber consists of a cover and a lower part, the lower part of the vacuum chamber having a shoulder for receiving the lower filter support. In addition, recesses for the gripper of the robot are provided in the side walls of the chamber in order that the filter plates or filter supports can be inserted and removed automatically. For the exact receiving and guidance of the filter supports or the collector support the lower part of the vacuum chamber has guide tabs having correcting bevels. The guide tabs preferably have two different bevel angles, the first bevel angle being about 30° and the second bevel angle being about from 0° to 2°. Furthermore, the guide edges of the wall with which the filter supports come into contact on insertion can be bevelled. Preferably the cover has a bevelled guide edge so that when the cover is put in place it is centred using the guide edge of the cover. The edge bevel angle is preferably 30°. The cover also has in the wall region recesses for the robot gripper and a supporting surface for the upper filter support.
The sealing material for the upper filter holder preferably has a hardness of about 20 Shore, the seal at the join between the cover and the lower part being formed by a combination of an O-ring and a resilient sealing strip, the O-ring providing a seal of about 60 Shore and the sealing strip of about 30 Shore. Sealing is als
Bloecker Helmut
Kauer Gerhard
Frommer Lawrence & Haug
Gesellschaft fuer Biotechnologische Forschung mbH (GBF)
Handy Dwayne K
Santucci Ronald R.
Warden Jill
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