Chemistry: molecular biology and microbiology – Virus or bacteriophage – except for viral vector or... – Recovery or purification
Patent
1998-12-21
2000-08-08
Salimi, Ali
Chemistry: molecular biology and microbiology
Virus or bacteriophage, except for viral vector or...
Recovery or purification
435181, 435820, 536 254, C12N 702, C12N 1106, C07H 2100
Patent
active
061000795
DESCRIPTION:
BRIEF SUMMARY
This application claims priority from PCT/JP97/00515, field Feb. 24, 1997.
FIELD OF THE INVENTION
The present invention relates to a method of processing biopolymers, microorganisms or substances, including DNA, RNA, mRNA, plasmid, viruses, bacteria and cells (all these are referred to as "DNA and the like") by using two or more kinds of magnetic particles, which enables automatic performance of processing, such as capture of cells, dissolution of cell nuclei and proteins, extraction and isolation of DNA and the like, and labeling, measuring or recovery of DNA and the like.
DESCRIPTION OF RELATED ART
In recent years, research on DNA and the like has been conducted in many fields such as engineering, medicine, agriculture, science and pharmacy, and its objectives range widely from genome sequencing, clinical diagnoses, breed improvement of agricultural plants, food bacillus inspection, and medicine development system.
Analysis of DNA and the like, whose application involves a wide range of fields and is expected to expand, has been performed in many ways, including centrifugal separation, high-performance liquid chromatography, gel electrophoresis, disposable column method, dialysis method, glass powder method, magnetic particle washing nozzle method, and immune serum reaction.
With the centrifugal separation method, however, loading and takeup of containers is very difficult to be automated and, after centrifugation, it is also very difficult to mechanically fractionate supernatants and precipitates. This method therefore lacks versatility.
With the high-performance liquid chromatography, because separation columns are essentially consumables, management of injection of samples into the columns and of separation time cannot be automated. Another problem with this method is that because samples are passed through the same column, contamination cannot be prevented completely.
In the case of the gel electrophoresis, adjustment of the gel cannot be automated and, although the DNA separation is a generally known as fundamental technique, there is no alternative but to use it manually to extract the separated fragments.
The disposable column method, one of methods that can be used as a kit to extract specific DNA fragments, is very costly and has a limited range of use. A further problem is the difficulty in controlling the pipetting of liquids and the amount of liquids supplied into columns, and many problems remain to be solved before automation can be realized.
The dialysis method takes time and has difficulty dealing with small amounts of samples and thus is not widely used.
The glass powder method is an excellent method of extracting DNA by using silicon dioxide. Although its process is simple, this method separates powder by filter or centrifugal separation and thus has a drawback of not being able to be automated easily.
The magnetic particle washing nozzle method, although it can be automated by controlling liquid suction and pouring by means of a cylinder and magnetic particles, has a problem that contamination basically cannot be eliminated by washing the nozzle.
The immune serum reaction is normally carried out by either a liquid phase method or a solid phase method. The liquid phase method has a similar problem to that experienced by the centrifugal separation. The solid phase method, too, has a problem similar to that of the centrifugal separation and requires many kinds of filters for separation from solid carriers, which makes the total automation difficult to realize. Another problem is that the use of many kinds of solid carriers makes this method difficult to handle and there is no appropriate means to avoid non-specific binding, giving rise to a limitation on highly sensitive and specific analyses.
The present invention has been accomplished under these situations and its objective is to first provide a method of processing DNA and the like, which, only by combining at least two kinds of magnetic particles, enables automatic and consistent performance of a series of processing including
Precision System Science Co., Ltd.
Salimi Ali
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