1. Field of the Invention
The present invention relates to a method of manufacturing a kit for isolating or purifying nucleic acids or biological materials from blood, cells or various biological samples, a kit manufactured by the method, and an apparatus using the kit.
2. Description of the Prior Art
Isolation or purification of nucleic acids from the blood or other various biological samples has been an important starting step in the fields of many areas such as biology, biochemistry, molecular medicine, forensic medicine, medical diagnostics, etc. Recently, a variety of Polymerase Chain Reaction (hereinafter, referred to as “PCR”) for DNA amplification (U.S. Pat. No. 4,683,195) have rendered the isolation of nucleic acids from biological samples to be more frequent and essential steps in both research and diagnostic areas. Conventional methods for isolating nucleic acids involve harmful organic solvents such as phenol and chloroform. (Sambrook, J., E. F. Fritsoh and T. Maniatis 1989, Molecular cloning: A Laboratory Manual, 2nd ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.)
Recently, several methods have been proposed using materials that have the proclivity of binding nucleic acids. Concrete examples of these materials are silica (Boom, R., Sol, C. J. A. Salimans, M. M. M., Jansen, C. L., Wertheim-van Dillen, P. M. E., and van der Noordaa, J. (1990) J. Clin. Microbiol. 28, 495-503), glass fibers (Vogelstein, B., and Gillespie, D. (1979) Proc. Natl. Acs. Sci. USA 76, 615-619; and Marko, M. A., Chipperfield, R., and Birnboim, H. C. (1982) Anal. Biochem. 121, 382-387), anion exchange resins (Wang, K., Boysen, C., and Hood, L. (1995) Anal. Biochem. 226, 85-90) and modified magnetic beads (Rudi, K., Kroken, M., Dahlberg, O. J., Deggerdal, A., Jakobsen, K. S., and Larsen, F. (1997) BioTechniques 22, 506-511; and Deggerdal, A. and Larsen, F. (1997) BioTechniques 22, 554-557).
The advantages of the methods using these materials are that no harmful organic solvents are involved, and that physical and biochemical degrading of nucleic acids during the isolation process is minimized. In addition, immobilized nucleic acids are less susceptible to digestion by nucleic acid-degrading enzymes such as nuclease.
The aforementioned methods, however, still need intensive manual pipetting steps to transfer the solid materials to other vessels, and thus, the performer is vulnerable to potential viral and bacterial infection from infectious viruses and bacteria if infected blood or bacteria is the starting material of nucleic acid isolation.
In order to avoid the intensive and tedious manual steps and to eliminate operator's potential error, several automatic machines such as “MagNa Pure LC” (Roche, Mannheim, Germany AG), “GENESIS” (Techan, Hombrechtikon, Switzerland) and others were developed for a large number of sample manipulations based on the concept disclosed in U.S. Pat. No. 3,985,649. Most of the automatic machines use magnetic beads to collect nucleic acids or biological materials from various biological samples and to eliminate the use of harmful chemical solvents and centrifugation steps. Although these machines are adequate for high throughput isolation of the nucleic acids or biological materials and secure the high throughput, they are also big, expensive, rather complicated, and inefficient for a small or medium number of sample manipulations. As a result, these machines are not practical for most diagnostic clinical and small research laboratories.
Furthermore, in order to operate these machines, the operator should from time to time supply solid materials such as magnetic beads and sufficient buffers to the machines. However, it is cumbersome and difficult for any person who has not trained for biological or clinical experiments to manage the machines.
In addition, the isolated nucleic acids obtained from the conventional machines are not convenient for the PCR amplification since they should be transferred to tubes containing the PCR-ready buffer.
In light of the drawbacks of the conventional machines for isolating nucleic acids or biological materials from the biological samples, there is still a need for a kit and an apparatus which are small, portable and efficiently applicable to the PCR amplification or other biological experiments.