US005656493A United States Patent (19 11 Patent Number: 5,656,493 Mullis et al. 45 Date of Patent: *Aug. 12, 1997

54 SYSTEM FOR AUTOMATED 4,582,789 4/1986 Sheldon, III et al...... 435/6 PERFORMANCE OF THE POLYMERASE 4,598,049 7/1986 Zelinka ...... 436/89 CHAIN REACTION 4,683,194 7/1987 Saiki et al...... 435/6 4,683,195 7/1987 Mullis et al...... 435/6 4,683.202 7/1987 Mullis ...... 435/9 75) Inventors: Kary B. Mullis, LaJolla; Larry 4,685,081 8/1987 Richman . ... 36.5/1 Johnson, San Jose; Richard A. Leath, 4,708,886 11/1987 Nelson ...... 422/72 Berkley; Timothy J. Wennberg, 4,711851 12/1987 McNamara. ... 422/63 Mariposa, all of Calif.; Louis M. 4,737,462 4/1988 Market al. . 435/253 Mezei, Madison, Wis.; Joseph T. 4,800,159 1/1989 Mullis et al. 435/1723 4853,332 8/1989 Market al...... 435/252.33 Widunas, Freemont, Calif. 4,858,155 8/1989 Okawa et al...... 422/63 4,865,186 9/1989 Coy et al...... 422/63 73) Assignee: The Perkin-Elmer Corporation, 4,865,986 9/1989 Coy et al...... 435/290 Norwalk, Conn. 4,889,818 12/1989 Gelfand et al...... 435/194 4,933,146 6/1990 Meyer et al...... 422/63 *) Notice: The term of this patent shall not extend 4,965,188 10/1990 Mullis et al...... 435/6 beyond the expiration date of Pat. No. 4,981,801 1/1991 Suzuki et al...... 435/290 5,038,852. 5,008,182 4/1991 Sninsky et al...... 435/5

5,038,852 8/1991 Johnson et al...... 165/12 5,075.216 12/1991 Innis et al...... 435/6 (21) Appl. No.: 199,505 5,333,675 8/1994 Mullis et al...... 165/12 22 Filed: Feb. 18, 1994 FOREIGN PATENT DOCUMENTS O110408 - 11/1983 European Pat. Off...... G05B 19/10 Related U.S. Application Data 171140 5/1984 European Pat. Off.. 63 Continuation of Ser. No. 21,624, Feb. 22, 1993, Pat No. 0 325 763 2/1989 European Pat. Off. . 5,333,675, which is a continuation of Ser. No. 709,374, Jun. 2413708 12/1977 France. 3, 1991, abandoned, which is a continuation of Ser. No. 2650593 5/1977 Germany ...... 357/87 899,061, Aug. 22, 1986, abandoned, which is a continuation 53-99.063 8/1978 Japan. in-part of Ser. No. 833,368, Feb. 25, 1986, abandoned, said 2161815 1/1986 United Kingdom. Ser. No. 709,374, is a continuation of Ser. No. 716,975, Mar. 28, 1985, abandoned. OTHER PUBLICATIONS (51] Int. Cl. ... C12M 3/02; C12M 1/00; "PC Application Ideas” Instruments and Control Systems G01N 21/00; C12N 15/00 (Oct. 1980). 52 U.S. Cl...... 435/286.1; 435/6: 435/91.1; 435/91.2: 435/287.2; 435/285.1; 422/50; Techne TP-16 Operating Manual May 1984. 422/62; 236/1 C; 236/15 RG; 236/91 R; Techne TP-16 Operating Manual Dec. 1984. 236/97; 935/76; 935/77; 935/87; 935/88 Smith, M., Appl. of Synthetic 58) Field of Search ...... 435/6, 183, 91.1, Oligodeoxyribonucleotides . . . pub. in Nucleic Acids Syn 435/91.2, 286.11, 287.1, 287.2, 285.1; 536/23.1, thesis: Applications to Molecule Biology and Genetic Engi 24.3, 24.33, 25.3; 935/76, 77, 85, 87,88; neering . . . Nucleic Acids Research-Nucleic Acids Sym 422/50, 62; 236/91 R, 97, 1 C, 15 BG posium Series No. 7, pp. 387-396 (1980). 56 References Cited (List continued on next page.) U.S. PATENT DOCUMENTS Primary Examiner-Bradley L. Sisson Attorney, Agent, or Firm-William J. Hone; Richard P. 3,311,303 3/1967 Noyes ...... 236/46 Ferrara 3,392,914 7/1968 Nienstaedt ... 236/46 3,856,471 12/1974 Winitz...... 23/253R 57) ABSTRACT 3,912,913 10/1975 Bunting ...... 235/150. 3,983,363 9/1976 Alter ...... 219/521 There is disclosed herein a machine for performing nucleic 4,008,048 2/1977 Hellemans .. ... 422/138 acid amplification under computer control. The machine 4206,872 6/1980 Levine ...... 236/46 4,312,835 l/1982 Zoltan et al...... 422/70 utilizes any one of a number of heating and cooling systems 4,335,620 6/1982 Adams ...... 73/863.11 under control of a host computer which directs the heating 4,358,535 11/1982 Falkow et al...... 435/5 and cooling systems to heat and cool a reaction-chamber 4,362,699 12/1982 Verlander et al...... 422/131 containing heat exchanger at appropriate times in the pro 4,395,486 7/1983 Wilson et al...... 435/6 cess. The reaction chambers are pre-loaded with the nucleic 4,404