Isolation of RNA from a mixture and its detection by utilizing a microgel-based optical device

Abstract

In this investigation, we show that RNA can be separated from a solution containing DNA and RNA and the isolated RNA can be detected using poly (N-isopropylacrylamide-co-N-(3-aminopropyl) methacrylamide hydrochloride) microgel-based optical devices (etalons). The isolation of RNA was accomplished by using hairpin-functionalized magnetic beads (MMPDNA) and differential melting, based on the fact that the DNA–RNA hybrid duplex is stronger (i.e., high melting temperature) than the DNA–DNA duplex (i.e., low melting temperature). By performing concurrent etalon sensing and fluorescent studies, we found that the MMPDNA combined with differential melting was capable of selectively separating RNA from DNA. This selective separation and simple colorimetric detection of RNA from a mixture will help lead to future RNA-based disease diagnostic devices.

Contrast variation SANS measurement of shell monomer density profiles of smart core–shell microgels

“…[1][2][3][4][5][6][7][8][9][10] This behaviour grants them the adjective smart. The response of these materials has great potential in the context of different applications like smart catalyst carriers, [11][12][13] drug delivery, 14,15 sensors, [16][17][18] optical devices, [19][20][21] responsive surface coatings 22 and actuators 23 or in biomedical applications. 24 Combining two monomers of different transition temperatures in statistical copolymers can be used to tune the swelling transition temperature, [25][26][27][28][29] whereas the subsequent synthesis of core and then shell 27,30-35 may lead to a continuous, linear temperature dependent swelling.…”

Surface‐Bound Microgels for Separation, Sensing, and Biomedical Applications

“…Glass is not an exclusively planar substrate, and substantial work has been dedicated to microgel-coated glass fibers to PNIPAm BSA Bioadhesion [67] PNIPAm and PEG Bioadhesion [45] PSS PDMAEMA PDDA Fundamental characterization [65] P(NIPAm-co-AA) Fundamental characterization [57] P(NIPAm-co-AA) PAH Fundamental characterization [201] PVA Alginate FGF Biomedical [163] PVDF P(AA-co-BA) Separations [101] PNIPAm Separations [205] P(NIPAm-co-AA) Separations [3] P4VP core PNIPAm shell Separations [72] Other Cotton P(NIPAm-co-AA) Separations [76] P(NIPAm-co-chitosan) Bioadhesion [102] PVCL Controlled release [19] Gold PNIPAm Fundamental characterization [89] PNIPAm AuNPs Sensing [9] PNIPAm Sensing [61] PNIPAm Sensing [108] P(NIPAm-co-APMAH) Sensing [43] P(NIPAm-co-APMAH) Sensing [106] P(NIPAm-co-APMAH) Sensing [107] P(NIPAm-co-AA) Sensing [11] P(NIPAm-co-AA) Sensing [33] P(NIPAm-co-AA) Sensing [41] P(NIPAm-co-AA) Sensing [62] P(NIPAm-co-AA) Sensing [110] P(NIPAm-co-AA) AuNPs, PDADMAC Fundamental characterization [150] P(NIPAm-co-AA) Ferrocene Sensing [12] P(NIPAm-co-AA) Phenyl boronic acid Sensing [88] P(NIPAm-co-AA) Phenyl boronic acid Sensing [90] P(NIPAm-co-AA) CV Controlled Release [147] P(NIPAm-co-GEMA) Sensing [10] P(Am-co-GMA) Phenyl boronic acid Sensing [13] P(NIPAm-co-4VP) Sensing [53] Graphite P(NIPAm-co-AA) carbon nanotubes Sensing [221] P(NIPAm-co-DMAPMA) butyrylcholinesterase Sensing [64] P(NIPAm-co-DMAPMA) choline oxidase Sensing [109]…”

“…[11] The authors further developed etalon sensors to detect DNA and RNA using P(NIPAm-co-(3-aminopropyl) methacrylamide hydrochloride) microgels, where the cationic microgels interacted with the negatively charged oligonucleotides, forming physical crosslinking that resulted in a blue shift of the reflectance spectrum proportional to the oligonucleotide concentration. [43,217] Zhang et al fabricated a peroxide-detecting sensor using P(NIPAm-co-AA) microgels modified with ferrocene. The microgel etalon contracted as the concentration of H 2 O 2 increased from 0.6 to 35 mm due to ferrocene oxidation and could be later reduced for reuse.…”

“…Many other measurement techniques have been developed for low concentration detection of pesticides, oxidants, and Glucose titer Glucose oxidase mediated redox reaction of glucose to gluconic acid Conductivity [62] Crosslinking density mediated reversible microgel swelling QCM [13] Deswelling of microgels induced by glucose binding to phenyl boronic acid Reflectance spectroscopy [88,90] Crosslinking density mediated reversible microgel swelling SPR [10] Peroxide titer Redox detection of myoglobin reducing H 2 O 2 Conductivity [110] Charge mediated reversible microgel swelling Reflectance spectroscopy [12] Crosslinking density mediated reversible microgel swelling Reflectance spectroscopy [223] Antioxidant titer Carbon nanotube mediated redox reaction Conductivity [221] Choline titer Enzymatic degradation of choline triggering redox detection Conductivity [109,215] DNA titer Charge mediated reversible microgel swelling Reflectance spectroscopy [43] IgG titer Charge mediated reversible microgel swelling Reflectance spectroscopy [106] Light Photothermal heating inducing LCST reversible microgel swelling Reflectance spectroscopy [9] Napthol-1 titer Nanoparticle arrangement controlled using microgel self-assembly to optimize SERS detection SPR [216] pH Charge mediated reversible microgel swelling Reflectance spectroscopy [41] Polycation titer Charge mediated reversible microgel swelling Reflectance spectroscopy [41] Progesterone titer Crosslinking density mediated reversible microgel swelling Reflectance spectroscopy [11] Pyrene titer Nanoparticle arrangement controlled using substrate patterning to optimize SERS detection SPR [180] Rhodamine 6G titer Nanoparticle arrangement controlled using substrate patterning to optimize SERS detection SPR [181] Temperature LCST mediated reversible microgel swelling Reflectance spectroscopy [105] Temperature/pH/salt titer LCST or charge mediated reversible microgel swelling QCM [33] www.afm-journal.de www.advancedsciencenews.com…”

“…Specifically, controlled swelling has been leveraged to release physically bound molecules, [20,[34][35][36] adjust the surface stiffness, [5,25,[37][38][39] and enable sensing activity. [9,40,41] Furthermore, the monomers used to construct the microgels are often amenable to post-synthetic modification with (bio)macromolecules, [42][43][44][45][46] nanoparticles, [47][48][49] and small molecules, [50][51][52][53] thus expanding their technological potential.…”