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Nallur, Girish; Marrero, R; Luo, Cheng-Hua; Krishna, R. Murli; Bechtel, Pamela E.; Shao, Weiping; Ray, Melissa; Wiltshire, Steve; Fang, Linhua; Huang, Hsing Hua; Liu, Chang–Gong; Sun, Lei; Sawyer, Jaymie R.; Schweitzer, Barry; Xia, Jing

doi:10.1023/a:1024535110995

Cited by 14 publications

(6 citation statements)

References 14 publications

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“…The concatamers are frequently thousands of nucleotides in length, leading to a large amplification of the initial circular “signal”. RCA has been an especially useful method for on-chip signal amplification because RCA-amplified concatamers can be localized to a given microarray spot. , …”

mentioning

confidence: 99%

Using a Deoxyribozyme Ligase and Rolling Circle Amplification To Detect a Non-nucleic Acid Analyte, ATP

et al. 2005

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An allosteric ribozyme (aptazyme) has been used to transduce the binding of a small organic analyte (ATP) into the ligation of a circular template for rolling circle amplification (RCA). An ATP-activated deoxyribozyme ligase was immobilized on a glass slide and, upon addition of ATP, catalyzed the ligation of a circular padlock probe. The ligated products could be directly amplified and visualized via RCA. The coupled reaction exhibited could detect as little as 1 muM of ATP and could discriminate against structurally similar nucleotides such as GTP, CTP, and UTP. Cooperative ATP activation of the deoxyribozyme was faithfully mimicked by RCA, yielding an amplified "switch" that was responsive to ATP concentration.

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mentioning

confidence: 99%

Using a Deoxyribozyme Ligase and Rolling Circle Amplification To Detect a Non-nucleic Acid Analyte, ATP

et al. 2005

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“…Recently developed spectrally encoded hydrogel beads provide an appealing alternative that have been used for a variety of biomedical and sensing applications 20,21 . Hydrogel beads are comprised of a cross-linked, hydrated polymeric network made up of one or more hydrophilic monomers, providing near fluid-phase kinetics at functionalized surfaces as well as high-efficiency molecular loading 22,23 , and microfluidic droplet generators have been used to produce and polymerize hydrogel beads at high-throughput 24 . However, the use of fluorescent dyes to create spectral codes in most cases has limited the number of unique spectral codes to < 100 25,26 .…”

Section: Introductionmentioning

confidence: 99%

MRBLES 2.0: High-throughput generation of chemically functionalized spectrally and magnetically-encoded hydrogel beads using a simple single-layer microfluidic device

Feng

White

Hein

et al. 2020

Preprint

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Widespread adoption of bead-based multiplexed bioassays requires the ability to easily synthesize encoded microspheres and conjugate analytes of interest to their surface. Here, we present a simple method (MRBLEs 2.0) for efficient high-throughput generation of microspheres with ratiometric barcode lanthanide encoding (MRBLEs) bearing functional groups for downstream bioconjugation. Bead production in MRBLEs 2.0 relies on manual mixing of lanthanide/polymer mixtures (each of which comprises a unique spectral code) followed by droplet generation using single-layer, parallel flow-focusing devices and off-chip batch polymerization of droplets into beads. To streamline downstream analyte coupling, MRBLEs 2.0 crosslinks copolymers bearing functional groups on the bead surface simultaneously during bead generation. Using the MRBLEs 2.0 pipeline, we generate monodisperse MRBLEs containing 48 distinct well-resolved spectral codes in high-throughput (>150,000/min and can be boosted to 450,000/min). We further demonstrate the efficient conjugation of oligonucleotides and entire proteins to carboxyl MRBLEs and biotin to amino MRBLEs. Finally, we show that MRBLEs can also be magnetized via simultaneous incorporation of magnetic nanoparticles with only a minor decrease in the potential code space. We anticipate that MRBLEs 2.0 can be directly applied towards a wide variety of downstream assays from basic biology to diagnostics and other translational research.

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“…Therefore, it is essential to develop methods to improve detection sensitivity and resolution such as nuclei amplification, which is an essential tool for many investigators to monitor infectious diseases, genetic disorders and traits. Polymerase chain reaction (PCR; Saiki et al 1985), self-sustained sequence replication (Guatelli et al 1990), nuclei acid sequencebased amplification (Compton 1991), strand displacement amplification (Walker 1992), rolling circle amplification (RCA; Lizardi et al 1998;Nallur et al 2003) and loopmediated isothermal amplification (LAMP; Notomi et al 2000) are examples of existing established DNA amplification methods. The most extensively used technique is PCR where the subsequent DNA synthesis is performed by heat denaturation of double-stranded DNA products.…”

Section: Introductionmentioning

confidence: 99%

Loop-mediated isothermal amplification of a single DNA molecule in polyacrylamide gel-based microchamber

Lam

Sakakihara

Ishizuka

et al. 2008

Biomed Microdevices

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Loop-mediated isothermal amplification (LAMP) is an original nucleic acid amplification method established by Notomi et al. LAMP is performed under isothermal condition, employing only a basic reaction protocol and minimal supporting electronics. These requirements prove to be viable for exploring the avenues to down-scale this biological reaction for Lab-on-a-chip application. Hence here, we developed a novel technique for fluorescent imaging of LAMP at a single molecule level. The experiment was conducted in a polyacrylamide (PAA) gel-based microchamber where a single DNA template, freely suspended in a solution containing primers and polymerase was initially encapsulated. In order to activate the amplification reaction, a microheater regulated by an automatic computerized feedback system was used for localized heating. This microchamber-based approach for LAMP demonstrated the effective exploitation of minute amount of templates and primers, and the overall reduction in LAMP detection time. An average efficiency of 80% was evaluated for conducting DNA amplification after 50 min of incubation at 65 degrees C. As the total time for reaction including detection can be completed in less than 1 h, this one-step, direct observation method displays the potential as a simple alternative to conventional techniques for genetic analysis and diagnosis in the clinical laboratory.

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Cited by 14 publications

References 14 publications

Using a Deoxyribozyme Ligase and Rolling Circle Amplification To Detect a Non-nucleic Acid Analyte, ATP

Using a Deoxyribozyme Ligase and Rolling Circle Amplification To Detect a Non-nucleic Acid Analyte, ATP

MRBLES 2.0: High-throughput generation of chemically functionalized spectrally and magnetically-encoded hydrogel beads using a simple single-layer microfluidic device

Loop-mediated isothermal amplification of a single DNA molecule in polyacrylamide gel-based microchamber

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