Abstract:
The present invention relates to a method for the diagnosis of tumoural conditions and/or of the corresponding state of advance, wherein a sample from a patient comprising at least one tumour cell is obtained. According to the invention, a purified specimen of the at least one tumour cell is obtained by individually selecting and isolating single cells in a microfluidic device the purified specimen having a purity of at least 90%. On the purified specimen thus obtained there is subsequently performed a molecular analysis such as to highlight a characteristic thereof suited to enabling diagnosis.
Abstract:
The present invention relates to a method for detecting a first and/or a second target DNA sequence from a DNA library, differing in that a mutation generates/eliminates a restriction site for a restriction endonuclease, comprising the steps of: (a) providing the DNA library, in which each of the DNA sequences comprises a first sequence segment, a second sequence segment of genomic DNA as cleaved by the restriction endonuclease, and a third sequence segment reverse complementary to the union of said first sequence segment and 5′ overhang, if any, of the restriction endonuclease; (b) amplifying the library of DNA sequences by PCR using: a first reverse primer which hybridizes to the 3′ end region of the second sequence segment of the first or second target sequence positive strand; a second forward primer which hybridizes to the 3′ end region of the second sequence segment of the first target sequence antipositive strand; a third forward primer comprising a first portion hybridizing to the 5′ end region of the third sequence segment of the second target sequence antipositive strand and a second portion hybridizing to the 3′ end region of the second sequence segment of the second target sequence antipositive strand, wherein the first portion of the third forward primer has a length from 20% to 80% with respect to the total length of the third forward primer; (c) detecting DNA sequences amplified in step (b).
Abstract:
Methods and apparatus are described for the processing (for example washing, incubation, etc.) of particles in which the particles suspended in a first fluid are introduced under laminar flow conditions into at least one first microchamber or first region of the same, in which a second fluid is introduced under laminar flow conditions into at least one second region of the microchamber or of a second microchamber, in such a way as not to mix with the first fluid, and in which at least one field of force acting on the particles is activated in the microchamber(s), to provoke a shift of the particles alone in a predetermined direction and to transfer the same in suspension into the second fluid; an apparatus is preferably used including at least three microchambers n microchambers arranged in sequence with each other in one direction and each connected with the microchamber immediately before it and after it with two orifices offset from each other in a direction perpendicular to the direction of sequence of the microchambers.
Abstract:
There is disclosed a method of generating a massively parallel sequencing library comprising the steps of: a) providing a primary WGA DNA library (pWGAlib), including fragments comprising a WGA library universal sequence adaptor; b) re-amplifying the primary WGA DNA library using at least one first primer (1PR) and at least one second primer (2PR); the at least one first primer (1PR) comprising from 5′ to 3′ at least one first sequencing adaptor (1PR5SA), at least one first sequencing barcode (1PR5BC) and a first primer 3′ section (1PR3S) hybridizing to either the WGA library universal sequence adaptor or its reverse complementary; the at least one second primer (2PR) comprising from 5′ to 3′ at least one second sequencing adaptor (2PR5SA) different from the at least one first sequencing adaptor (1PR5SA), and a second primer 3′ section (2PR3S) hybridizing to either the WGA library universal sequence adaptor or its reverse complementary.
Abstract:
A microfluidic device for the isolation of particles of at least one specific type of a sample is described. The microfluidic device comprises: a first inlet adapted to receive a sample comprising the particles of the specific type, at least part of a separation group comprising a separation unit, which comprises a main chamber and a recovery chamber and being adapted to receive the sample and to transfer at least part of the particles of the specific type from the main chamber to the recovery chamber in a selective manner with respect to further particles of the sample, a first outlet which is designed to allow the particles of the specific type to be collected outside of the device and a second outlet adapted to allow at least a portion of the sample to flow out of the main chamber and out of the microfluidic device.
Abstract:
Method for the treatment of a biological sample comprising at least one cell and a liquid component; according to the method, a force is applied to the sample inserted in an inner chamber of a hollow device towards a filter which has pores with diameters from 2 nm to 1 μm, so that at least part of the liquid component passes through the filter and the cell remains in the inner chamber, thus obtaining a concentrated sample; the filter has a surface facing the inner chamber of less than 12.6 mm2.
Abstract:
Methods and apparatus are described for the processing particles in which the particles suspended in a first fluid are introduced under laminar flow conditions into at least one first microchamber or first region, in which a second fluid is introduced under laminar flow conditions into at least one second region or second microchamber, in such a way as not to mix with the first fluid, and in which at least one field of force acting on the particles is activated in the microchamber(s), to provoke a shift of the particles alone in a predetermined direction and to transfer the same in suspension into the second fluid.
Abstract:
The present disclosure relates to a method for analyzing the degree of similarity of at least two samples in a plurality of samples comprising genomic DNA. The method comprises the following steps. a) Providing a plurality of samples comprising genomic DNA. b) Carrying out, separately on each sample, a deterministic restriction-site whole genome amplification (DRS-WGA) of said genomic DNA, c) Preparing a massively parallel sequencing library using a fragmentation-free, sequencing-adaptor/WGA fusion-primer PCR reaction from each product of said DRS-WGA. d) Carrying out low-pass whole genome sequencing at a mean coverage depth of
Abstract:
Method and device (1b) for performing the optical analysis of particles (2) contained in suspension in a fluid (3) arranged inside a microfluidic device (4) which maintains it at a temperature significantly lower than the ambient temperature; the formation of humidity on the outer surface (8) of the cover of the microfluidic device is avoided by applying a thermal flow (P) which determines an increase in the temperature of the outer surface (8) of the cover to above the condensation temperature (Td), or reduction in the ambient temperature (and/or humidity) in the vicinity of the cover (8), so as to bring the condensation temperature (Td) (dew point) to below the temperature of the surface (8) of the cover determined by the internal operating temperature.
Abstract:
There is disclosed a method of generating a massively parallel sequencing library comprising the steps of: a) providing a primary WGA DNA library (pWGAlib), including fragments comprising a WGA library universal sequence adapter; b) performing a single PCR cycle on the pWGAlib using a first primer (1PR) comprising from 5′ to 3′ a first sequencing adapter (1PR5SA) and a first primer 3′ section (1PR3S) hybridizing to the reverse complementary of the WGA library universal sequence adapter; c) performing a single PCR cycle on the on the product of step b) using a second primer (2PR) comprising from 5′ to 3′ a second sequencing adapter (2PR5SA) different from the 1PR5SA, and a second primer 3′ section (2PR3S) hybridizing to the WGA library universal sequence adapter reverse complementary; d) amplifying by PCR the product of step c) using a third primer comprising the 1PR5SA and a fourth primer comprising 2PR5SA.