Journal: Advanced healthcare materials

Article Title: Woven Electroanalytical Biosensor for Nucleic Acid Amplification Tests.

doi: 10.1002/adhm.202100034

Figure Lengend Snippet: Figure 4. Schematics of E-DNA sensor fabrication, integration into the Coolmax-weave, and isothermal amplification RPA. A) Schematics of immobi- lization of methylene blue conjugated stem-loop (S-L) DNA probes on the clean gold microwire surface and later backfilling of the electrode surface by 6MCH. B) Schematics of the mechanism of recombinase polymerase amplification (RPA) and providing ssDNA target using lambda exonuclease enzyme for the detection step. (C) and (D) schematics of the woven E-DNA sensor into Coolmax-cotton weave, including a stem-loop (S-L) DNA probe modified-gold microwire, a silver microwire, and double-gold microwires as working (WE), pseudo-reference (RE), and counter (CE) electrodes, in ab- sence and presence of RPA target sample. E) Gel electrophoresis image of RPA amplified S. epidermidis genomic DNA with a target size of 210 bp, before converting to single-stranded DNA using exonuclease lambda enzyme as dsDNA product and after digesting the untargeted strand as ssDNA. F) Photo of the real E-DNA sensor integrated into the woven microfluidic devices.

Article Snippet: Lambda exonuclease enzyme, phosphate-buffered saline (PBS) tablets pH 7.4, nuclease-free water, and 10× Tris-borate-EDTA (TBE) buffer were purchased from Fisher Scientific (Sweden).

Techniques: Nucleic Acid Electrophoresis

Journal: Advanced healthcare materials

Article Title: Woven Electroanalytical Biosensor for Nucleic Acid Amplification Tests.

doi: 10.1002/adhm.202100034

Figure Lengend Snippet: Figure 5. Detecting RPA products through woven E-DNA sensors into the Coolmax-cotton weave using signal suppression (SS = (ΔIb −ΔIss) / ΔIb)), in which ΔIb is the baseline peak current and ΔISS is the suppressed peak current. A) Typical SWV scans of the woven E-DNA sensor for added 60 µL of 1× PBS (pH 7.4) after 5 min. (B) Typical SWV scans of the woven E-DNA sensor for added 60 µL of unpurified 250-fold diluted RPA blank (treated by lambda exonuclease enzyme) product as a negative control sample after 5 min of hybridization. C) Typical SWV scans of the woven E-DNA sensor for added 60 µL of unpurified 250-fold diluted ssDNA RPA target product after 5 min of hybridization. D) Signal suppression of the integrated E-DNA sensors into the woven devices in presence of the unpurified RPA amplified ssDNA target (with an initial 10 copies/µL of S. epidermidis genomic DNA), the RPA blank product treated with lambda enzyme and PBS (1×, pH 7.4) after 5 min of hybridization. Reported data are based on the independent woven E-DNA sensors (n = 4) for RPA products and (n = 6) for PBS (1×, pH 7.4). Statistical analysis is carried out using Student’s t-test (two-tailed) and ** p < 0.05 is considered significant.

Article Snippet: Lambda exonuclease enzyme, phosphate-buffered saline (PBS) tablets pH 7.4, nuclease-free water, and 10× Tris-borate-EDTA (TBE) buffer were purchased from Fisher Scientific (Sweden).

Techniques: Negative Control, Hybridization, Two Tailed Test