Author: Lifeng Zhou; Arun Richard Chandrasekaran; Jibin Abraham Punnoose; Gaston Bonenfant; Stephon Charles; Oksana Levchenko; Pheonah Badu; Cassandra Cavaliere; Cara T. Pager; Ken Halvorsen
Title: Programmable low-cost DNA-based platform for viral RNA detection Document date: 2020_1_16
ID: 8kced06y_19
Snippet: Oligonucleotides were purchased from Integrated DNA Technologies (IDT) with standard desalting, and the full sequences of all strands is listed in Supporting information Table S1 to S9. Nanoswitches were constructed as described previously. 24, 27 A genomic single-stranded DNA (New England Biolabs M13mp18) was linearized using targeted cleavage with BtsCI restriction enzyme. The linearized ssDNA was then mixed with a molar excess of an oligonucle.....
Document: Oligonucleotides were purchased from Integrated DNA Technologies (IDT) with standard desalting, and the full sequences of all strands is listed in Supporting information Table S1 to S9. Nanoswitches were constructed as described previously. 24, 27 A genomic single-stranded DNA (New England Biolabs M13mp18) was linearized using targeted cleavage with BtsCI restriction enzyme. The linearized ssDNA was then mixed with a molar excess of an oligonucleotide mixture containing backbone oligos and detectors, and annealed from 90°C to 20°C at 1 °C min −1 in a T100™ Thermal Cycler (Bio-Rad, USA). Following construction, the nanoswitches were purified using liquid chromatography (LC) purification 29 to remove excess oligonucleotides. The concentration of purified nanoswitches were determined by measuring A260 absorbance with a Thermo Scientific NanoDrop 2000.
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