Detailed design information and probe sequences are listed in the (Additional file 1 : Table S1). ssDNAs were purchased from Integrated DNA Technologies (Coralville, IA). Probes were generated by hybridization of equimolar amounts of complementary ssDNAs in 1× STE buffer (10 mM Tris, 100 mM NaCl, and 1 mM EDTA), heating to 95 °C for 5 min, followed by incubation at room temperature for 1 h. After hybridization, dsDNA probes were 5′-radiolabeled with 10 pmoles of [γ-33P] ATP using T4 polynucleotide kinase (New England Biolabs, Ipswich, MA). Radiolabeled DNA probes were purified from unincorporated label with G-25 Sephadex columns (Roche Applied Science; Indianapolis, IN).
Ssdnas
Manufactured by Integrated DNA Technologies
Single-stranded DNA (ssDNA) is a type of DNA molecule that consists of a single strand of nucleotides. ssDNA is commonly used as a starting material in various molecular biology applications, such as DNA sequencing, PCR amplification, and probe synthesis. The core function of ssDNA is to provide a template for downstream processes and analyses.
Automatically generated - may contain errors
Lab products found in correlation
G 25 sephadex column, by Roche (1 mentions)
T4 polynucleotide kinase, by New England Biolabs (1 mentions)
Titanium 4 isopropoxide, by Thermo Fisher Scientific (1 mentions)
Dimethyl sulfoxide (dmso), by Thermo Fisher Scientific (1 mentions)
Dithiothreitol (dtt), by Thermo Fisher Scientific (1 mentions)
Sodium chloride (nacl), by Thermo Fisher Scientific (1 mentions)
Hydrochloric acid (hcl), by Thermo Fisher Scientific (1 mentions)
Mgcl2, by Thermo Fisher Scientific (1 mentions)
Ovalbumin (ova), by Merck Group (1 mentions)
Tris base, by Thermo Fisher Scientific (1 mentions)
Cytochrome c, by Merck Group (1 mentions)
Polyvinylpyrrolidone, by Merck Group (1 mentions)
Ethanol etoh, by Thermo Fisher Scientific (1 mentions)
Guanidine hcl gu hcl, by Thermo Fisher Scientific (1 mentions)
25 mm magnesium chloride mgcl2 solution, by New England Biolabs (1 mentions)
Hemoglobin, by Merck Group (1 mentions)
Taq polymerase, by New England Biolabs (1 mentions)
β casein, by Merck Group (1 mentions)
Glacial acetic acid, by Thermo Fisher Scientific (1 mentions)
2 protocols using ssdnas
1
Radiolabeling of DNA Probes
2
Aptamer Selection and Protein Binding Assays
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Dithiothreitol (DTT), NaCl, MgCl2, 1× PBS buffer, glacial acetic acid, hydrochloric acid, DMSO, titanium (IV) isopropoxide, guanidine HCl (Gu-HCl) and potassium chloride (KCl) were obtained from Fisher Scientific. Polyvinylpyrrolidone (PVP) was acquired from Sigma-Aldrich. Tris base, EDTA and ethanol (EtOH) was obtained from Acros Organics, Promega and Decon Labs, respectively. The 5× Taq polymerase and the 25-mM magnesium chloride (MgCl2) solution used for PCR were from New England Biolabs. The ssDNAs, including the random libraries used for SELEX, the anti-IgE aptamer, as well as those found binding to DNMT1 (Supplementary Tables S1 and S3 ) were attained from Integrated DNA Technologies, Inc. Beta-casein, cytochrome C, ovalbumin, and hemoglobin were purchased from Sigma-Aldrich. IgE was acquired from Athens Research & Technology, Inc.
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