Method, kit and apparatus for the isolation of nucleic acids
OwnerJorg Kleiber, Christine Markert-Hahn +1inventors
InventorsJorg Kleiber, Christine Markert-Hahn, Herbert Harttig
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Abstract
The invention concerns a process for preparing biological samples for the subsequent detection of an analyte. In particular, the invention relates to a process for the isolation of a nucleic acid in a sample using a suspension of magnetic glass particles. In addition, kits and apparatuses containing magnetic glass particles for sample preparation are provided.
Claims43
Process for the isolation of an analyte from a biological sample comprising the steps:
- (a) lysing the sample in a reaction vessel,
- (b) adding a solid adsorption matrix
- (c) incubating under such conditions that the analyte binds to the adsorption matrix,
- (d) removing non-bound sample components from the reaction vessel,
- (e) incubating under such conditions that the analyte is eluted from the adsorption matrix and
- (f) separating the eluate from the adsorption matrix.
Process as claimed in one of the previous claims,
- wherein
- magnetic glass particles are used as the solid adsorption matrix.
Process as claimed in one of the previous claims,
- wherein
- the amount of adsorption matrix added is at most 50% more than the amount that is required to quantitatively bind the analyte present in the sample.
Process as claimed in one of the previous claims,
- wherein
- a continuous or intermittent mixing without adding external devices is carried out at least during steps (c), (d) or/and (e).
Process as claimed in one of the previous claims,
- wherein
- step (d) comprises adding and aspirating a wash buffer which is optionally repeated several times.
Process as claimed in one of the previous claims,
- wherein
- additional reagents such as enzymes are added in step (e).
Process as claimed in one of the previous claims,
- wherein
- a low salt buffer is used for elution in step (e).
Process as claimed in one of the previous claims,
- wherein
- at least steps (c) and (d) are carried out at essentially the same temperature.
Process as claimed in one of the previous claims,
- wherein
- step (a) or/and step (e) are carried out at an elevated temperature.
Process as claimed in one of the previous claims,
- wherein
- an aftertreatment step at an elevated temperature is carried out after step (f).
Process for the isolation of an analyte from a biological sample comprising the steps:
- (a) lysing the sample in a reaction vessel,
- (b) adding a solid adsorption matrix
- (c) incubating under such conditions that the analyte binds to the adsorption matrix,
- (d) separating non-bound sample components from the adsorption matrix,
- (e) incubating under such conditions that the analyte is eluted from the adsorption matrix and
- (f) separating the eluate from the adsorption matrix
- wherein at least steps (c) and (d) are carried out at essentially the same temperature.
Process as claimed in one of the previous claims,
- wherein
- the analyte is a nucleic acid.
Process as claimed in one of the previous claims,
- wherein
- the process is carried out in an automated device.
Reagent kit for the isolation of DNA comprising magnetic glass particles whose glass phase contains SiO2, B2O3 and Na2O.
Reagent kit for the isolation of RNA comprising magnetic glass particles whose glass phase contains SiO2, B2O3, Al2O3, CaO and K2O.
Device for isolating an analyte from a biological sample comprising:
- a sample preparation device (1),
- a holding device for reagents (2),
- a first holding device for reaction vessels for sample preparation (3) which is equipped for an operating temperature of ≦70° C., in particular ≦40° C.,
- a second holding device for reaction vessels (4a, 4b, 4c), which optionally contains a cooling or/and heating means,
- and a robotic tool device (5).
Magnetic glass particles comprising a magnetic core and a glass coat which contains SiO2, B2O3, an alkali metal oxide and optionally Al2O3 and an alkaline earth metal oxide.
Dependencies are inferred from the claim text.