Recombinant DNA screening system including fixed array replicator and support

• US 4,591,567 A
• Filed: 08/08/1983
• Issued: 05/27/1986
• Est. Priority Date: 04/21/1982
• Status: Expired due to Fees

• Current Assignee: California Institute of Technology
• Original Assignee: California Institute of Technology
• Inventors: Davidson, Eric H., Britten, Roy J.
• Primary Examiner(s): Tanenholtz, Alvin E.
• Assistant Examiner(s): NOT, DEFINED
• Application Number:
  US06/520,756
• Time in Patent Office: 1,023 Days
• Field of Search: 435/5, 435/6, 435/30, 435/35, 435/172, 435/292, 435/293, 435/294, 435/800, 435/809, 935/80, 935/85, 935/86
• US Class Current: 435/285.1
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: C12M 23/50 Means for positioning or orientating the apparatus C12M41/08 takes precedence
• CPC Class Codes: Y10S 435/80 Elimination or reduction of contamination by undersired ferments, e.g. aseptic cultivation
• CPC Class Codes: Y10S 435/809 Incubators or racks or holders for culture plates or containers

Abstract

A system for the production and maintenance of genomic libraries of recombinant DNA in a fixed geometric array arranged so that such libraries may be accurately replicated and maintained with the same set of individual DNA inserts occupying reproducible locations and comprising,

(a) sets of individual bacterial colonies or plaques of bacteriophage which contain inserts of animal or plant DNA and are present in so large a number that there will be a high probability that the complete library will contain each DNA sequence of all of the sequences present in an animal or plant genome; to be established in subarrays which can individually be replicated,

(b) a subsystem for picking up inocula from each of a geometric array simultaneously and transferring them to a second similar geometric array, including carrying and aligning means for maintaining a transfer means in superposed relationship with respect and for raising and lowering the transfer means into and out of proximity to said colonies or plaques, and

(c) a subsystem for intially establishing such subarrays by transfer from a randomly spread pattern of colonies or plaques of bacteriophage.

Claims

1. A device adapted to replicate recombinant DNA libraries of eukaryotic genomes in fixed arrays comprising:

(a) container means of a size adapted to contain phage or bacteria in a random array of at least about 50,000 plaques,

(b) in proximity to said container means, a multipoint simultaneous transfer means containing at least about 50,000 rigid points having fixed distance between points and being adapted to transfer phage or bacteria from colonies such that locations are precisely maintained, all of the free ends of said points terminating a plane for simultaneous contact with and penetration of the surface containing the phage or colony,

(c) carrying and aligning means for maintaining said transfer means in superposed relationship with respect to said container means including vertical slide members adjacent corners of said transfer means whereby said transfer means can be slidably moved toward and away from said container means while maintaining the container means and transfer means in essentially constant registry, said container means and transfer means being generally coextensive in area and shape, and said container means being provided with micrometer adjustment and restraining means in both the "X" and "Y" axes for controlled small lateral displacement of said container means with respect to said transfer means and said carrying and aligning means, and

(d) a base member, wherein said vertical slide members, said container means and said adjustment and restraining means are carried by said base member.

2. The device of claim 1 wherein the multipoint simultaneous transfer means is a flat support to which a large number of pins are affixed.

3. The device of claim 2 wherein the pins are spaced uniformly from each other at a spacing of about 2 millimeters.

4. The device of claim 2 wherein the pins are in the form of crimped wires and said free ends are the apexes of said crimped wires, and wherein the pins are affixed to the support with epoxy resin.

5. The device of claim 1 wherein the multipoint simultaneous transfer means is a flat support having a plurality of identical pyramids projecting upwardly therefrom said pyramids being uniformly spaced.

6. The device of claim 5 wherein the pyramids are spaced at about 1/10 of an inch at their apexes, said pyramids and the support are integrally formed, and the apexes of one row or pyramids are staggered with respect to the apexes of the pyramids of immediately adjoining rows.