and were in vitro transcribed to generate synthetic RNA targets. In vitro transcription was conducted as previously described22. In brief, the dsDNA template was transcribed using the HiScribe T7 high-yield RNA synthesis kit (New England Biolabs (NEB)) ...
happened as in Fuet al.50. Captured samples as well as raw libraries for shotgun sequencing were indexed using universal iP5 and unique iP7 indexes49. NGS sequencing was performed on an Illumina MiSeq System using the Illumina MiSeq Reagent Kit v3 (150-cycles). AmpFLSTR Yfiler PCR Amplifica...
MinElute PCR Purification Kit QIAGEN Cat# 28006 TwistAmp Basic Kit TwistDX Cat# TABAS03kit Qubit dsDNA HS Assay Kit, 500 assays Thermo Fisher Scientific Cat# Q32854 High Pure Extender Assembly from the Roche High Pure Viral Nucleic Acid Large Volume Kit,40 reactions Roche Cat# 5114403001 MiSe...
The following parameters were optimized using the TwistAmp Basic Kit: Reaction temperature and incubation time: RPA amplification reactions were conducted at temperatures ranging from 37 to 41 °C and varying incubation times between 5 and 20 min. These parameters were systematically varied to determine...
The RPA reaction was performed using 3 µL of RNA and a TwistAmp® RT Basic kit (TwistDx, Cambridge, UK) for 25 min at 40 ◦C. RPA products were analyzed on a 2% agarose gel and by lateral flow assay (LFA) using a Milenia HybriDetect 1 kit (TwistDx). Clinical samples of TB...
A diagnostic recombinase polymerase amplification (RPA) assay for Aphelenchoides fragariae using a TwistAmp Basic Kit (TwistDx, Cambridge, UK) and AmplifyRP Acceler8 Discovery Kit (Agdia, Elkhart, IN, USA) combined with lateral flow dipsticks (LF) has been developed. In this study, a LF-RPA ...
Multiplex RPA Reactions in Solution RPA was performed according to the manufacturer's instructions with a TwistAmp Basic Kit (TwistDX, Cambridge, UK). Briefly, the reaction mixtures contained 25 µL 2× reaction buffer; 11.5 µL ddH2O; 2 µL each of Staphylococcus aureus, Vibrio ...
The commercial TwistAmp® Exo kit (TwsitDx, Cambridge, UK) was used for real-time RPA assays, following the manufacturer's instructions. Lyophilised RPA proteins were reconstituted with rehydration solution, forward and reverse primers, a fluorescent exo-probe and DNA samples containing plasmids ...