Organic compounds -- part of the class 532-570 series – Organic compounds – Carbohydrates or derivatives
Reexamination Certificate
1999-08-27
2001-05-15
Tsang, Cecilia (Department: 1623)
Organic compounds -- part of the class 532-570 series
Organic compounds
Carbohydrates or derivatives
C536S025410, C536S025420, C422S069000, C422S070000, C210S450000, C210S462000, C210S465000, C210S600000, C210S613000, C210S615000, C204S602000
Reexamination Certificate
active
06232464
ABSTRACT:
BACKGROUND OF THE INVENTION
The invention relates to a process and an apparatus for isolating nucleic acids.
Before the analysis by polymerase chain reaction (PCR) of nucleic acids obtained from cells, it is necessary to purify and concentrate the nucleic acids. In addition, it can be necessary to remove from the sample to be analyzed certain substances interfering with the polymerase chain reaction, such as the hemoglobin prosthetic group.
In addition, in other techniques of nucleic acid analysis, for example hybridization, purification and concentration of the nucleic acids to be analyzed also plays an important role.
“Methods of Enzymology”, Vol. 68, pp. 170-182, discloses using “spin-columns” for the isolation of nucleic acids. In a variant of this technique disclosed by DE 41 39 664 A1, the following working steps are used:
aa) cell disintegration,
bb) adsorption of the nucleic acids to a glass fiber fleece in the presence of a high ionic strength buffer, and
cc) elution of the nucleic acids by a low ionic strength buffer.
In the step aa), the liquid sample is passed through a glass fiber fleece to which the nucleic acids adsorb. The glass fiber fleece is then washed with various solutions. Finally, the nucleic acids are eluted from the solid phase in the presence of low ionic strength buffers.
The known process is disadvantageous in a number of respects: contamination of the sample can occur during washing of the solid phase. In addition, because of the capillary forces prevailing in the glass fiber fleece, the nucleic acids can only be partially recovered therefrom.
Furthermore, “Methods in Enzymology 65” (1980), pp. 371-380 discloses gel electrophoretic methods in which nucleic acids are bound to gels and are then brought back into solution by electroelution. Contamination of the solution can also occur in this case. The nucleic acids are present in the solution at high dilution. Concentration does not occur.
The object of the present invention is to specify a process and an apparatus by which the disadvantages of the prior art are avoided. In particular, a process and an apparatus for isolating nucleic acids which enable simple and inexpensive purification and concentration of nucleic acids are to be specified. Furthermore, a substantially automated isolation and concentration of nucleic acids is to be able to be carried out. Finally, the purpose of the invention is to avoid contaminations.
SUMMARY OF THE INVENTION
According to the invention, a process is provided for isolating nucleic acids from biological fluids and suspensions containing nucleic acids,
a) the nucleic acids being bound to an adsorption medium,
b) the nucleic acids being eluted from the adsorption medium, and
c) being moved by electrophoresis from a reaction compartment into a removal compartment connected thereto and enriched there.
The process makes possible in a simple manner purification and concentration of nucleic acids from liquids. In particular in an automatic process procedure, the risk of contamination can be largely excluded. The elution can be performed by buffer change or electrically by electroelution or electrophoresis.
Further according to the invention, an apparatus for isolating nucleic acids from biological fluids and suspensions containing nucleic acids is provided, a reaction compartment for receiving an adsorption medium laden with nucleic acids being connected to a removal compartment, and the nucleic acids being able to be moved by an electrophoresis device from the reaction compartment into the removal compartment and enriched there. This apparatus makes a simple and inexpensive concentration and isolation of nucleic acids possible. By the provision of a separate removal compartment, contamination can be largely excluded.
Exemplary embodiments of the invention are described in more detail below with reference to the drawing.
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Tsuda, “Chromatographic Behavior in Electrochromatography,”Anal Chem.,60(17), 1677-1680 (Sep. 1, 1988).
Crane L. Eric
Tsang Cecilia
Vanophem Meehan & Vanophem P.C.
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