Ruggedized apparatus for analysis of nucleic acid and proteins

The invention provides methods and systems for ruggedizing a nucleic acid analyzing apparatus. The ruggedized apparatus can be used reliably and effectively in uncontrolled environments, such as, for example at a crime scene to collect and analyze forensic data, as well as in semi-controlled environments, such as, for example at a point of care location.

Methods for Rapid Multiplexed Amplification of Target Nucleic Acids

A fast, multiplexed PCR system is described that can rapidly generate amplified nucleic acid products, for example, a full STR profile, from a target nucleic acid. Such systems include, for example, microfluidic biochips and a custom built thermal cycler, which are also described. The resulting STR profiles can satisfy forensic guidelines for signal strength, inter-loci peak height balance, heterozygous peak height ratio, incomplete non-template nucleotide addition, and stutter. 1. A thermal cycler comprising:a temperature control element (TCE), wherein a first surface of said TCE is adapted to receive a sample chamber containing a solution and a sensing chamber containing a thermosensor,wherein said thermosensor provides feedback to said TCE to set or maintain the solution at a desired temperature.2. A thermal cycler comprising:a temperature control element (TCE), wherein a first surface of said TCE is adapted to receive a sample chamber containing a solution and a sensing chamber containing a first thermosensor,wherein said thermosensor provides feedback to said TCE to set or maintain the solution at a desired temperature, further comprising a second thermosensor positioned to monitor the temperature of said first surface of said TCE.3. A thermal cycler comprising:a temperature control element (TCE), wherein a first surface of said TCE is adapted to receive a sample chamber, said chamber containing a solution and a thermosensor, wherein said thermosensor provides feedback to said TCE to set or maintain the solution at a desired temperature.4. The thermal cycler of claim 2 , further comprising a chip compression element (CCE) positioned over the first surface of the TCE allowing insertion of a substrate between the CCE and the TCE claim 2 , wherein the CCE provides for thermal communication between the TCE and the substrate.5. The thermal cycler of claim 4 , wherein the CCE comprises a low thermal mass and insulating material.6. The thermal cycler of claim 4 , ...

Plastic Microfluidic Separation and Detection Platforms

Plastic electrophoresis separation chips are provided comprising a plurality of microfluidic channels and a detection window, where the detection window comprises a thin plastic; and the detection window comprises a detection region of each microfluidic channel. Such chips can be bonded to a support provided an aperture is provided in the support to allow detection of samples in the electrophoresis chip at the thin plastic detection window. Further, methods for electrophoretically separating and detecting a plurality of samples on the plastic electrophoresis separation chip are described. 1. A plastic electrophoresis chip for separation and detection of DNA fragments comprising:a substrate having top and bottom surfaces, further comprising an anode portion, a cathode portion, and a center portion between the anode and cathode portions, anda cover layer having top and bottom surfaces such that the top surface of said substrate layer is bonded with the bottom surface of said cover layer to form a plurality of non-intersecting microfluidic channels and a detection window, each nonintersecting microfluidic channel having an equivalent length of between 2-50 cm and a separation and detection region; and the length, widths and depths of each of the plurality of microfluidic channels is adjusted so that the resistances in each of the plurality of microfluidic channels is substantially equivalent and configured to detect DNA; and wherein,each non-intersecting microfluidic channel is in fluid communication with a first via located at said anode anda second via located at said cathode,said detection window comprised of plastic selected from the group consisting of polyethylene, poly(carbonate) and poly(acrylate) which essentially do not fluoresce light having a wavelength between 500 and 800 nm when excited at a wavelength between about 450 and 500 nm, and said detection window has a thickness of less than 2 mm, and overlaps in the detection region of each of said plurality ...

Integrated Nucleic Acid Analysis

The present disclosure provides fully integrated microfluidic systems to perform nucleic acid analysis. These processes include sample collection, nucleic acid extraction and purification, amplification, sequencing, and separation and detection. The present disclosure also provides optical detection systems and methods for separation and detection of biological molecules. In particular, the various aspects of the invention enable the simultaneous separation and detection of a plurality of biological molecules, typically fluorescent dye-labeled nucleic acids, within one or a plurality of microfluidic chambers or channels. The nucleic acids can be labeled with at least 6 dyes, each having a unique peak emission wavelength. The present systems and methods are particularly useful for DNA fragment sizing applications such as human identification by genetic fingerprinting and DNA sequencing applications such as clinical diagnostics. 1. An optical detector comprisingone or more light sources positioned for illuminating one or a plurality of detection positions on a substrate, said substrate comprising a thermal cycling chamber adapted for multiplexed amplification in one PCR reaction, such that primers labeled with 6 or more fluorescent dyes each having a unique peak emission wavelength are used to generate amplified nucleic acid fragments labeled with said 6 or more fluorescent dyes, said thermal cycling chamber in fluid communication with at least one channel on said substrate, wherein each channel comprises a detection position;a mirror to scan said one or more light sources sequentially between detection positions;one or a plurality of first optical elements positioned for collecting and directing light emanating from the detection positions on the substrate; anda light detector positioned to accept light directed from the one or plurality of first optical elements, wherein the light detector comprises a wavelength dispersive element to disperse the light from the one ...

Integrated systems for the multiplexed amplification and detection of six and greater dye labeled fragments

The present disclosure provides fully integrated microfluidic systems to perform nucleic acid analysis. These processes include sample collection, nucleic acid extraction and purification, amplification, sequencing, and separation and detection. The present disclosure also provides optical detection systems and methods for separation and detection of biological molecules. In particular, the various aspects of the invention enable the simultaneous separation and detection of a plurality of biological molecules, typically fluorescent dye-labeled nucleic acids, within one or a plurality of microfluidic chambers or channels. The nucleic acids can be labeled with at least 6 dyes, each having a unique peak emission wavelength. The present systems and methods are particularly useful for DNA fragment sizing applications such as human identification by genetic fingerprinting and DNA sequencing applications such as clinical diagnostics.

Methods for rapid multiplexed amplification of target nucleic acids

A fast, multiplexed PCR system is described that can rapidly generate amplified nucleic acid products, for example, a full STR profile, from a target nucleic acid. Such systems include, for example, microfluidic biochips and a custom built thermal cycler, which are also described. The resulting STR profiles can satisfy forensic guidelines for signal strength, inter-loci peak height balance, heterozygous peak height ratio, incomplete non-template nucleotide addition, and stutter.

Ruggedized Apparatus for Analysis of Nucleic Acid and Proteins

The invention provides methods and systems for ruggedizing a nucleic acid analyzing apparatus. The ruggedized apparatus can be used reliably and effectively in uncontrolled environments, such as, for example at a crime scene to collect and analyze forensic data, as well as in semi-controlled environments, such as, for example at a point of care location.

Plastic Microfluidic Separation and Detection Platforms

Plastic electrophoresis separation chips are provided comprising a plurality of microfluidic channels and a detection window, where the detection window comprises a thin plastic; and the detection window comprises a detection region of each microfluidic channel. Such chips can be bonded to a support provided an aperture is provided in the support to allow detection of samples in the electrophoresis chip at the thin plastic detection window. Further, methods for electrophoretically separating and detecting a plurality of samples on the plastic electrophoresis separation chip are described.

Integrated Nucleic Acid Analysis

The present disclosure provides fully integrated microfluidic systems to perform nucleic acid analysis. These processes include sample collection, nucleic acid extraction and purification, amplification, sequencing, and separation and detection. The present disclosure also provides optical detection systems and methods for separation and detection of biological molecules. In particular, the various aspects of the invention enable the simultaneous separation and detection of a plurality of biological molecules, typically fluorescent dye-labeled nucleic acids, within one or a plurality of microfluidic chambers or channels. The nucleic acids can be labeled with at least 6 dyes, each having a unique peak emission wavelength. The present systems and methods are particularly useful for DNA fragment sizing applications such as human identification by genetic fingerprinting and DNA sequencing applications such as clinical diagnostics.

Ruggedized Apparatus for Analysis of Nucleic Acid and Proteins

The invention provides methods and systems for ruggedizing a nucleic acid analyzing apparatus. The ruggedized apparatus can be used reliably and effectively in uncontrolled environments, such as, for example at a crime scene to collect and analyze forensic data, as well as in semi-controlled environments, such as, for example at a point of care location.

Ruggedized Apparatus for Analysis of Nucleic Acid and Proteins

The invention provides methods and systems for ruggedizing a nucleic acid analyzing apparatus. The ruggedized apparatus can be used reliably and effectively in uncontrolled environments, such as, for example at a crime scene to collect and analyze forensic data, as well as in semi-controlled environments, such as, for example at a point of care location. 1. A system for finding the position of separation channels in an electrophoresis system comprising an electrophoresis chip and an instrument capable of separating and detecting of DNA fragments labeled with at least one fluorescent dye comprising:(a) a removable electrophoresis chip comprising:a substrate having top and bottom surfaces, further comprising an anode portion, a cathode portion, and a separation channel between the anode and cathode portions, andhaving top and bottom surfaces such that the top surface of said substrate layer is bonded with the bottom surface of said cover layer to form at least one microfluidic channel,a first via located at said anode and in fluidic communication with at least one of the microfluidic channels,a second via located at said cathode and in fluidic communication with at least one of the microfluidic channels,a detection zone located in the separation channel; and(b) an electrophoresis instrument capable of receiving said chip, comprising,(i) a processor,(ii) an optical system, said optical system comprising said fluorescence excitation energy beam, and at least one photodiode, positioned on a first side of said chip when said chip is inserted into said instrument, and at least one light detector, and,(iii) an electrical system for delivering said beam to said detection zone, and having an anode and a cathode, said anode in contact with said anode portion of said substrate, and said cathode in contact with said cathode portion of said substrate, andwhereby the processor directs the beam to pass through the detection zone, and the scattered light from the separation channel ...

METHODS FOR RAPID MULTIPLEXED AMPLIFICATION OF TARGET NUCLEIC ACIDS

A fast, multiplexed PCR system is described that can rapidly generate amplified nucleic acid products, for example, a full STR profile, from a target nucleic acid. Such systems include, for example, microfluidic biochips and a custom built thermal cycler, which are also described. The resulting STR profiles can satisfy forensic guidelines for signal strength, inter-loci peak height balance, heterozygous peak height ratio, incomplete non-template nucleotide addition, and stutter. 169- (canceled)70. A method for simultaneously amplifying of a plurality of loci in a nucleic acid solution comprising:providing in a single solutions contained in at least two reaction chambers located in a biochip, samples having at least ten target nucleic acid loci to be amplified, with at least ten different primer pairs, each primer pair hybridizing to one of the at least ten loci to be amplified, said solution further comprising:(i) one or more buffers;(ii) one or more salts;(iii) a nucleic acid polymerase; and(iv) nucleotides; andproviding a thermal control system comprised of (i) a TCE having a first surface in thermal communication with said at least two reaction chambers, said TCE further comprising a means for heating and cooling, (ii) at least one thermosensor, (iii) a controller that receives signals from said at least one thermosensor and (iv)a power supply, said at least one thermosensor positioned and configured to measure the effective temperature of each of the single solutions in the reaction chambers of the biochip and to provide feedback to the TCE to heat or cool the solution to set or maintain the solution at a desired temperature;obtaining, with said controller of the thermal control system, a target sample solution temperature, from a control algorithm that stores the target sample temperature, for a sample for a first sub-step of a plurality of predetermined processing sub-steps, to be performed on the samples, where in the first sub-step of the processing step is ...

Integrated Nucleic Acid Analysis

The present disclosure provides fully integrated microfluidic systems to perform nucleic acid analysis. These processes include sample collection, nucleic acid extraction and purification, amplification, sequencing, and separation and detection. The present disclosure also provides optical detection systems and methods for separation and detection of biological molecules. In particular, the various aspects of the invention enable the simultaneous separation and detection of a plurality of biological molecules, typically fluorescent dye-labeled nucleic acids, within one or a plurality of microfluidic chambers or channels. The nucleic acids can be labeled with at least 6 dyes, each having a unique peak emission wavelength. The present systems and methods are particularly useful for DNA fragment sizing applications such as human identification by genetic fingerprinting and DNA sequencing applications such as clinical diagnostics. 1. A method for multiple-sample DNA analysis , comprising:injecting a first template DNA extracted based on a first sample from a first sample chamber on a biochip through a first channel to a first reaction reservoir in a first region of said biochip;injecting a second template DNA extracted based on a second sample from a second sample chamber on said biochip through a second inlet to a second reaction reservoir in the first region of said biochip, the second sample chamber being separate from the first sample chamber;inducing thermal cycles in the first region of the biochip for PCR amplification of DNA fragments, the first region including at least the first reaction reservoir designated for PCR amplification based on the first sample, and the second reaction reservoir designated for PCR amplification based on the second sample;inducing liquid flow to respectively move first amplified DNA fragments from the first reaction reservoir to a first separation unit in a second region of the biochip, and second amplified DNA fragments from the ...

Plastic Microfluidic Separation and Detection Platforms

Plastic electrophoresis separation chips are provided comprising a plurality of microfluidic channels and a detection window, where the detection window comprises a thin plastic; and the detection window comprises a detection region of each microfluidic channel. Such chips can be bonded to a support provided an aperture is provided in the support to allow detection of samples in the electrophoresis chip at the thin plastic detection window. Further, methods for electrophoretically separating and detecting a plurality of samples on the plastic electrophoresis separation chip are described. 129-. (canceled)30. A plastic chip component , comprising:(a) a separation chip having a substrate layer and a cover layer;wherein one of the substrate layer and cover layer includes a plurality of grooves and the other of the substrate layer or the cover layer includes a series of through holes;wherein a top surface of the substrate layer and a bottom surface of the cover layer are bonded together to form (i) a first microfluidic channel that connects the sample to one or a plurality of second microfluidic channels; and (ii) said one or a plurality of second microfluidic channels, each of said second microfluidic channels having a detection region at the termini of a separation region;wherein each of the substrate layer and the cover layer comprises a thin plastic film of polyethylene, a poly(carbonate), an unsaturated, partially unsaturated or saturated cyclic olefin polymer (COP), an unsaturated, partially unsaturated, or saturated cyclic olefin copolymer (COC), or a norbornene thermopolymer, or a poly (acrylate) which essentially does not fluoresce light having a wavelength between 500 and 800 when excited at a wavelength between about 450 and 500 nm,and wherein each of the substrate layer and the cover layer independently has a thickness of less than 200 μm; and(b) a plastic support fixedly attached to the chip havingan anode portion with at least one anode well,a cathode ...

Ruggedized apparatus for analysis of nucleic acid and proteins

The invention provides methods and systems for ruggedizing a nucleic acid analyzing apparatus. The ruggedized apparatus can be used reliably and effectively in uncontrolled environments, such as, for example at a crime scene to collect and analyze forensic data, as well as in semi-controlled environments, such as, for example at a point of care location.

Plastic microfluidic separation and detection platforms

Plastic electrophoresis separation chips are provided comprising a plurality of microfluidic channels and a detection window, where the detection window comprises a thin plastic; and the detection window comprises a detection region of each microfluidic channel. Such chips can be bonded to a support provided an aperture is provided in the support to allow detection of samples in the electrophoresis chip at the thin plastic detection window. Further, methods for electrophoretically separating and detecting a plurality of samples on the plastic electrophoresis separation chip are described.

Plastic microfluidic separation and detection platforms

Plastic electrophoresis separation chips are provided comprising a plurality of microfluidic channels and a detection window, where the detection window comprises a thin plastic; and the detection window comprises a detection region of each microfluidic channel. Such chips can be bonded to a support provided an aperture is provided in the support to allow detection of samples in the electrophoresis chip at the thin plastic detection window. Further, methods for electrophoretically separating and detecting a plurality of samples on the plastic electrophoresis separation chip are described.

Ruggedized apparatus for analysis of nucleic acid and proteins

The invention provides methods and systems for ruggedizing a nucleic acid analyzing apparatus. The ruggedized apparatus can be used reliably and effectively in uncontrolled environments, such as, for example at a crime scene to collect and analyze forensic data, as well as in semi-controlled environments, such as, for example at a point of care location.

Plastic microfluidic separation and detection platforms

Plastic electrophoresis separation chips are provided comprising a plurality of microfluidic channels and a detection window, where the detection window comprises a thin plastic; and the detection window comprises a detection region of each microfluidic channel. Such chips can be bonded to a support provided an aperture is provided in the support to allow detection of samples in the electrophoresis chip at the thin plastic detection window. Further, methods for electrophoretically separating and detecting a plurality of samples on the plastic electrophoresis separation chip are described.

Integrated nucleic acid analysis

Abstract The present disclosure provides fully integrated microfluidic systems to perform nucleic acid analysis. These processes include sample collection, nucleic acid extraction and purification, amplification, sequencing, and separation and detection. The present disclosure also provides optical detection systems and methods for separation and detection of biological molecules. In particular, the various aspects of the invention enable the simultaneous separation and detection of a plurality of biological molecules, typically fluorescent dye-labelled nucleic acids, within one or a plurality of microfluidic chambers or channels. The nucleic acids can be labelled with at least 6 dyes, each having a unique peak emission wavelength. The present systems and methods are particularly useful for DNA fragment sizing applications such as human identification by genetic fingerprinting and DNA sequencing applications such as clinical diagnostics. ?0l1 O0 3 6 7v1 106 107 104 165 108 FIGuR E I

Integrated nucleic acid analysis

The present disclosure provides fully integrated microfluidic systems to perform nucleic acid analysis. These processes include sample collection, nucleic acid extraction and purification, amplification, sequencing, and separation and detection. The present disclosure also provides optical detection systems and methods for separation and detection of biological molecules. In particular, the various aspects of the invention enable the simultaneous separation and detection of a plurality of biological molecules, typically fluorescent dye-labeled nucleic acids, within one or a plurality of microfluidic chambers or channels. The nucleic acids can be labeled with at least 6 dyes, each having a unique peak emission wavelength. The present systems and methods are particularly useful for DNA fragment sizing applications such as human identification by genetic fingerprinting and DNA sequencing applications such as clinical diagnostics.

Ruggedized apparatus for analysis of nucleic acid and proteins

The invention provides methods and systems for ruggedizing a nucleic acid analyzing apparatus. The ruggedized apparatus can be used reliably and effectively in uncontrolled environments, such as, for example at a crime scene to collect and analyze forensic data, as well as in semi-controlled environments, such as, for example at a point of care location.

Plastic microfluidic separation and detection platforms

Plastic electrophoresis separation chips are provided comprising a plurality of microfluidic channels and a detection window, where the detection window comprises a thin plastic; and the detection window comprises a detection region of each microfluidic channel. Such chips can be bonded to a support provided an aperture is provided in the support to allow detection of samples in the electrophoresis chip at the thin plastic detection window. Further, methods for electrophoretically separating and detecting a plurality of samples on the plastic electrophoresis separation chip are described.

Plastic microfluidic separation and detection platforms

Plastic microfluidic separation and detection platforms

Plastic electrophoresis separation chips are provided comprising a plurality of microfluidic channels and a detection window, where the detection window comprises a thin plastic; and the detection window comprises a detection region of each microfluidic channel. Such chips can be bonded to a support provided an aperture is provided in the support to allow detection of samples in the electrophoresis chip at the thin plastic detection window. Further, methods for electrophoretically separating and detecting a plurality of samples on the plastic electrophoresis separation chip are described.

Plastic microfluidic separation and detection platforms

Plastic electrophoresis separation chips are provided comprising a plurality of microfluidic channels and a detection window, where the detection window comprises a thin plastic; and the detection window comprises a detection region of each microfluidic channel. Such chips can be bonded to a support provided an aperture is provided in the support to allow detection of samples in the electrophoresis chip at the thin plastic detection window. Further, methods for electrophoretically separating and detecting a plurality of samples on the plastic electrophoresis separation chip are described.

Methods for rapid multiplexed amplification of target nucleic acids

Plastic microfluidic separation and detection platforms

Ruggedized apparatus for analysis of nucleic acid and proteins

Integrated systems for the multiplexed amplification and detection of six and greater dye labeled fragments

The present disclosure provides fully integrated microfluidic systems to perform nucleic acid analysis. These processes include sample collection, nucleic acid extraction and purification, amplification, sequencing, and separation and detection. The present disclosure also provides optical detection systems and methods for separation and detection of biological molecules. In particular, the various aspects of the invention enable the simultaneous separation and detection of a plurality of biological molecules, typically fluorescent dye-labeled nucleic acids, within one or a plurality of microfluidic chambers or channels. The nucleic acids can be labeled with at least 6 dyes, each having a unique peak emission wavelength. The present systems and methods are particularly useful for DNA fragment sizing applications such as human identification by genetic fingerprinting and DNA sequencing applications such as clinical diagnostics.

Integrated microfluidic systems, biochips and methods for the detection of nucleic acids and biological molecules by electrophoresis

本揭示案提供完全整合微流體系統以進行核酸分析。此等過程包括樣品收集、核酸萃取與純化、擴增、測序及分離與偵測。本揭示案亦提供用於分離及偵測生物分子之光學偵測系統及方法。詳言之，本發明之各態樣使在一或複數個微流體室或通道內同時分離及偵測複數個生物分子(通常為經螢光染料標記之核酸)能夠進行。該等核酸可用至少6種染料標記，各染料具有獨特之峰值發射波長。本發明之系統及方法尤其適用於測DNA片段大小之應用(諸如藉由遺傳指紋進行人類識別)及DNA測序應用(諸如臨床診斷)。

Ruggedized apparatus for analysis of nucleic acid and proteins

The invention provides methods and systems for ruggedizing a nucleic acid analyzing apparatus. The ruggedized apparatus can be used reliably and effectively in uncontrolled environments, such as, for example at a crime scene to collect and analyze forensic data, as well as in semi-controlled environments, such as, for example at a point of care location.

Methods for rapid multiplexed amplification of target nucleic acids

A fast, multiplexed PCR system is described that can rapidly generate amplified nucleic acid products, for example, a full STR profile, from a target nucleic acid. Such systems include, for example, microfluidic biochips and a custom built thermal cycler, which are also described. The resulting STR profiles can satisfy forensic guidelines for signal strength, inter-loci peak height balance, heterozygous peak height ratio, incomplete non-template nucleotide addition, and stutter.

Plastic microfluidic separation and detection platforms

Plastic electrophoresis separation chips are provided comprising a plurality of microfluidic channels and a detection window, where the detection window comprises a thin plastic; and the detection window comprises a detection region of each microfluidic channel. Such chips can be bonded to a support provided an aperture is provided in the support to allow detection of samples in the electrophoresis chip at the thin plastic detection window. Further, methods for electrophoretically separating and detecting a plurality of samples on the plastic electrophoresis separation chip are described. Plastic electrophoresis separation chips are provided comprising a plurality of microfluidic channels and a detection window, where the detection window comprises a thin plastic; and the detection window comprises a detection region of each microfluidic channel. Such chips can be bonded to a support provided an aperture is provided in the support to allow detection of samples in the electrophoresis chip at the thin plastic detection window. Further, methods for electrophoretically separating and detecting a plurality of samples on the plastic electrophoresis separation chip are described.

Plastik mikroakışkan ayırma ve saptama platformları.

Plastik elektroforez ayırma çipleri, saptama penceresinin ince bir plastik içerdiği ve saptama penceresinin her bir mikroakışkan kanalın bir saptama bölgesini içerdiği çok sayıda mikroakışkan kanal ve bir saptama penceresinden oluşmaktadır. Bu gibi çipler, ince plastik saptama penceresindeki elektroforez çipindeki numunelerin saptanmasını sağlamak için destekte bir aralığın temin edilmesi şartıyla bir desteğe bağlanabilmektedir. Ayrıca, plastik elektroforez ayırma çipi üzerinde çok sayıda numunenin elektroforetik olarak ayırma ve saptama yöntemleri tarif edilmektedir.

Plastic microfluidic separation and detection platforms

Plastic electrophoresis separation chips are provided comprising a plurality of microfluidic channels and a detection window, where the detection window comprises a thin plastic; and the detection window comprises a detection region of each microfluidic channel. Such chips can be bonded to a support provided an aperture is provided in the support to allow detection of samples in the electrophoresis chip at the thin plastic detection window. Further, methods for electrophoretically separating and detecting a plurality of samples on the plastic electrophoresis separation chip are described.

Ruggedized apparatus for analysis of nucleic acid and proteins

The invention provides methods and systems for ruggedizing a nucleic acid analyzing apparatus. The ruggedized apparatus can be used reliably and effectively in uncontrolled environments, such as, for example at a crime scene to collect and analyze forensic data, as well as in semi-controlled environments, such as, for example at a point of care location.