Oligonucleotides were prepared on an Applied Biosystems Inc. 394 DNA synthesizer. Commercially available DNA synthesis reagents were obtained from Glen Research Inc. DNA substrates used in this study are presented in Figure 3 . T4 polynucleotide kinase, Klenow exo–, terminal deoxynucleotide transferase, and T4 DNA ligase were obtained from New England Biolabs. [α-32P]dCTP, [α-32P]-dTTP, [γ-32P]ATP, and [α-32P]cordycepin 5′-triphosphate were purchased from PerkinElmer. Analyses of radiolabeled oligonucleotides were conducted using a Storm 840 Phosphor-imager and ImageQuant TL software. C18-Sep-Pak cartridges were obtained from Waters.
Dna synthesis reagents
Manufactured by Glen Research
Sourced in United States
DNA synthesis reagents are a set of chemical compounds used in the process of synthesizing DNA molecules. These reagents include the necessary building blocks, such as nucleotides, and other essential components required for the controlled and efficient creation of DNA sequences.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
γ 32p atp, by PerkinElmer (2 mentions)
Terminal deoxynucleotidetransferase, by New England Biolabs (1 mentions)
Sep pak c18 cartridge, by Waters Corporation (1 mentions)
α 32p dttp, by PerkinElmer (1 mentions)
α 32p cordycepin 5 triphosphate, by PerkinElmer (1 mentions)
394 dna synthesizer, by Thermo Fisher Scientific (1 mentions)
α 32p dctp, by PerkinElmer (1 mentions)
Klenow exo, by New England Biolabs (1 mentions)
T4 polynucleotide kinase, by New England Biolabs (1 mentions)
T4 dna ligase, by New England Biolabs (1 mentions)
Milli q ultrapure water system, by Merck Group (1 mentions)
C18 sep pak cartridgeswere, by Waters Corporation (1 mentions)
Autoflex 3, by Bruker (1 mentions)
Ammonium persulfate, by Merck Group (1 mentions)
Acrylamide, by Merck Group (1 mentions)
4 protocols using dna synthesis reagents
1
Oligonucleotide Synthesis and Radiolabeling
2
DNA and Cell Culture Reagents
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All reagents were used as received from commercial sources or prepared as described in references. All DNA synthesis reagents were purchased from Glen Research (Sterling, VA). Protected dFe phosphoramidites were synthesized in our lab.
Dulbecco’s phosphate-buffered saline (DPBS), Dulbecco’s modified Eagles medium (DMEM), Leibovitz’s L-15 medium (L-15), and fetal bovine serum (FBS) were obtained from Thermo Fisher Scientific Inc. All solutions used in the experiments were prepared using ultrapure water (resistance > 18 MΩ cm), which was obtained through a Millipore Milli-Q ultrapure water system (Billerica, MA, USA).
Dulbecco’s phosphate-buffered saline (DPBS), Dulbecco’s modified Eagles medium (DMEM), Leibovitz’s L-15 medium (L-15), and fetal bovine serum (FBS) were obtained from Thermo Fisher Scientific Inc. All solutions used in the experiments were prepared using ultrapure water (resistance > 18 MΩ cm), which was obtained through a Millipore Milli-Q ultrapure water system (Billerica, MA, USA).
3
Oligonucleotide Synthesis and Characterization
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Oligonucleotides were synthesized via
standard automated DNA synthesis on an Applied Biosystems Inc. model
394 instrument. DNA synthesis reagents were obtained from Glen Research.
Oligonucleotides were purified by 20% denaturing gel electrophoresis
and desalted using C18-Sep-Pak cartridges. Oligonucleotides were characterized
by ESI (Thermoquest LCQ Deca) or MALDI-TOF (Bruker Autoflex III) mass
spectrometry.
Radiolabeling was carried out using standard protocols
and is briefly described below.34 T4 polynucleotide
kinase (PNK) was purchased from New England Biolabs. γ-32P-ATP was purchased form PerkinElmer. C18-Sep-Pak cartridges
were obtained from Waters. Quantification of radiolabeled oligonucleotides
was carried out using a Molecular Dynamics phosphorimager equipped
with ImageQuant TL software. Radiolabeled samples were counted using
a Beckman Coulter LS 6500 scintillation counter. Photolyses were carried
out in a Rayonet RPR-100 photoreactor (Southern New England Ultraviolet)
equipped with 16 lamps with maximum emission at 350 nm. BME and piperidine
solutions were freshly prepared.
standard automated DNA synthesis on an Applied Biosystems Inc. model
394 instrument. DNA synthesis reagents were obtained from Glen Research.
Oligonucleotides were purified by 20% denaturing gel electrophoresis
and desalted using C18-Sep-Pak cartridges. Oligonucleotides were characterized
by ESI (Thermoquest LCQ Deca) or MALDI-TOF (Bruker Autoflex III) mass
spectrometry.
Radiolabeling was carried out using standard protocols
and is briefly described below.34 T4 polynucleotide
kinase (PNK) was purchased from New England Biolabs. γ-32P-ATP was purchased form PerkinElmer. C18-Sep-Pak cartridges
were obtained from Waters. Quantification of radiolabeled oligonucleotides
was carried out using a Molecular Dynamics phosphorimager equipped
with ImageQuant TL software. Radiolabeled samples were counted using
a Beckman Coulter LS 6500 scintillation counter. Photolyses were carried
out in a Rayonet RPR-100 photoreactor (Southern New England Ultraviolet)
equipped with 16 lamps with maximum emission at 350 nm. BME and piperidine
solutions were freshly prepared.
4
Cocaine Nanopore Sensing Methodology
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Cocaine was obtained from National Institutes for Food and Drug Control (Beijing, China). Ammonium persulfate (APS), tetramethylethylenediamine (TEMED), and acrylamide were purchased from Sigma-Aldrich (St. Louis, MO, USA). 2-Cyanoethyl diisopropyl chlorophosphoramidite was purchased from Chem Genes (Wilmington, MA, USA). FC-40 (a mixture of perfluoro-tri-n-butylamine and perfluoro-di-n-butylmethylamine) was purchased from Minnesota Mining and Manufacturing Company (St. Paul, MN, USA). DNA synthesis reagents were purchased from Glen Research (Sterling, VA, USA). Other reagents were purchased from Sinopharm Chemical Reagent (Shanghai, China). The cocaine buffer contained 77 mM Na2HPO4, 23 mM NaH2PO4, 50 mM NaCl, 5 mM MgCl2 (pH 7.3). All solutions were prepared with ultra-pure Milli-Q water (resistance > 18 MΩ cm−1). Glass capillaries from Suzhou City Crystal Glass Co., Ltd. were used. The inner radius of these capillaries is constant and equal to 300 ± 5 μm.
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