1 H and 13C NMR spectra were measured with Varian NMR spectrometers (Inova 300, Varian 400, and Mercury 400). For optical spectra, DNA-CAs were prepared as ∼1 μM solutions in water (Molecular Biology grade, Corning). Absorption spectra were measured with a Cary 100 Bio UV–vis spectrometer, fluorescence spectra obtained by a Jobin Yvon-Spex Fluorolog 3 spectrometer, and fluorescence lifetime measurements were made with a PTI EasyLife LS spectrometer. Mass spectra were obtained using ESI or MALDI-TOF ionization modes at the Stanford University Mass Spectrometry Facility. HPLC was performed with a Shimadzu LC-20AD equipped with an SPD-M20A diode array detector and a Phenomenex Jupiter reverse phase C5 column. Dynamic light scattering measurements were made with a Malvern Instruments Nanoseries ZS90 Zetasizer. Epifluorescence microscopy for cell bioimaging and library screening were conducted on a Nikon Eclipse 80i microscope equipped with a Nikon Plan Fluor 4–40× objective and a QIClick digital CCD camera, with a 100 W high-pressure mercury lamp as the excitation source (365 nm mercury plasma emission line), 340–380 nm excitation filter, and >420 nm long-pass emission filter. Photographs of aqueous phase DNA-CAs, DNA-CAs in hydrogel, and flexible DNA-CA display were captured with an iPhone 6S+ camera, with a 365 nm gel transilluminator UV source (VWR LM-20E) as backlight.
Inova 300
Manufactured by Agilent Technologies
The Inova 300 is a nuclear magnetic resonance (NMR) spectrometer manufactured by Agilent Technologies. It is designed to analyze the structure and composition of chemical compounds through the detection and measurement of nuclear magnetic resonances.
Automatically generated - may contain errors
Lab products found in correlation
1200 series, by Merck Group (3 mentions)
1100 series, by Merck Group (3 mentions)
Chromolith rp 18e, by Merck Group (3 mentions)
Diode array detector, by Agilent Technologies (2 mentions)
Nmr spectrometer, by Agilent Technologies (1 mentions)
Mercury 400, by Agilent Technologies (1 mentions)
Eclipse 80i microscope, by Nikon (1 mentions)
Lc 20ad, by Shimadzu (1 mentions)
Spd m20a, by Phenomenex (1 mentions)
Acquity uplc, by Waters Corporation (1 mentions)
Cary 300, by Agilent Technologies (1 mentions)
Silica gel 60, by Merck Group (1 mentions)
Microtof q 2, by Bruker (1 mentions)
Heleos 2, by Wyatt Technology (1 mentions)
6130 series, by Merck Group (1 mentions)
Nano zs zen 3600, by Malvern Panalytical (1 mentions)
14 protocols using inova 300
1
Characterization of DNA-Conjugated Agents
2
Synthesis and Characterization of OGR1 Probe
3
Polycation Characterization by SEC-MALS
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The number average molecular
weight (Mn) and dispersity (Mw/Mn) for each polycation
was determined by size exclusion chromatography (SEC) equipped with
Eprogen columns [CATSEC1000 (7 μm, 50 × 4.6), CATSEC100
(5 μm, 250 × 4.6), CATSEC300 (5 μm, 250 × 4.6),
and CATSEC1000 (7 μm, 250 × 4.6)], a Wyatt HELEOS II light
scattering detector (λ = 662 nm), and an Optilab rEX refractometer
(λ = 658 nm). The columns were maintained at 30 °C. An
aqueous eluent (0.1 M Na2SO4/1 v/v % acetic
acid) was utilized at a flow rate of 0.4 mL/min. The dn/dc values for each of the polymers were determined
offline with the Optilab rEX refractometer.
1H NMR
measurements were performed with a Varian Inova 300 at 70 °C.
Samples were dissolved in D2O (HOD used as the internal
standard), and the block copolymer compositions were determined by
calculating the ratio between the integrals of resonances of the PMAG
block and those of the PAEMA or PDMAPMA block.
weight (Mn) and dispersity (Mw/Mn) for each polycation
was determined by size exclusion chromatography (SEC) equipped with
Eprogen columns [CATSEC1000 (7 μm, 50 × 4.6), CATSEC100
(5 μm, 250 × 4.6), CATSEC300 (5 μm, 250 × 4.6),
and CATSEC1000 (7 μm, 250 × 4.6)], a Wyatt HELEOS II light
scattering detector (λ = 662 nm), and an Optilab rEX refractometer
(λ = 658 nm). The columns were maintained at 30 °C. An
aqueous eluent (0.1 M Na2SO4/1 v/v % acetic
acid) was utilized at a flow rate of 0.4 mL/min. The dn/dc values for each of the polymers were determined
offline with the Optilab rEX refractometer.
1H NMR
measurements were performed with a Varian Inova 300 at 70 °C.
Samples were dissolved in D2O (HOD used as the internal
standard), and the block copolymer compositions were determined by
calculating the ratio between the integrals of resonances of the PMAG
block and those of the PAEMA or PDMAPMA block.
4
Synthesis and Characterization of SpiroZin1
5
Synthesis of Triblock Copolymer PLGA-PEG-PLGA
6
Non-invasive Intracellular pH Monitoring
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M. smegmatis was grown
to a cell density of 108 cells/mL in a total volume of
500 mL in a 4 L Erlenmeyer
flask with constant shaking at 37 °C in Difco Middlebrook 7H9
media supplemented with oleic acid/albumin/dextrose and 0.05% Tween
80. Cells were harvested by centrifugation, and the pellet was washed
twice with 5 mM phosphate buffer, pH 6.8. The cell pellet was then
resuspended in 200 μL of the same buffer and 500 μL of
the resulting cell slurry transferred to a 5 mm NMR tube. Chemical
shifts were referenced with respect to 85% phosphoric acid in D2O in a coaxial capillary. 31P NMR spectra were
obtained using a Varian INOVA 300 (at 121.5 MHz) using 60° pulse
excitation, proton decoupling, and a 1 s recycle time. A total of
1024 scans were accumulated corresponding to approximately a 60 min
total data acquisition time (without aeration). Spectra were analyzed
as described elsewhere.46 (link) The peak corresponding
to the α-phosphate of ATP (at ∼−10.5 ppm) and
the inorganic phosphate peaks of interest (in the region of 0–1.5
ppm) were used to calculate the internal and external pH using the
following equation, where d is the distance between
the α-phosphate of ATP and the inorganic phosphate peak, in
ppm.
to a cell density of 108 cells/mL in a total volume of
500 mL in a 4 L Erlenmeyer
flask with constant shaking at 37 °C in Difco Middlebrook 7H9
media supplemented with oleic acid/albumin/dextrose and 0.05% Tween
80. Cells were harvested by centrifugation, and the pellet was washed
twice with 5 mM phosphate buffer, pH 6.8. The cell pellet was then
resuspended in 200 μL of the same buffer and 500 μL of
the resulting cell slurry transferred to a 5 mm NMR tube. Chemical
shifts were referenced with respect to 85% phosphoric acid in D2O in a coaxial capillary. 31P NMR spectra were
obtained using a Varian INOVA 300 (at 121.5 MHz) using 60° pulse
excitation, proton decoupling, and a 1 s recycle time. A total of
1024 scans were accumulated corresponding to approximately a 60 min
total data acquisition time (without aeration). Spectra were analyzed
as described elsewhere.46 (link) The peak corresponding
to the α-phosphate of ATP (at ∼−10.5 ppm) and
the inorganic phosphate peaks of interest (in the region of 0–1.5
ppm) were used to calculate the internal and external pH using the
following equation, where d is the distance between
the α-phosphate of ATP and the inorganic phosphate peak, in
ppm.
7
NMR Spectroscopy and HPLC Analysis
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Example 4
General Methods:
1H NMR spectra were recorded on a Varian Inova 300 or 500 MHz NMR instrument. Chromatographic purities were determined on an Agilent 1200 Series, 1100 Series or 6130 Series LC/MS system using a Merck Chromolith RP-18e analytical HPLC column (monolithic, 50×2 mm) and the following analytical HPLC method: injection volume 5 μL; flow rate 1 mL/min; 5→95% acetonitrile in water with 0.05% AcOH over 5 mins; Agilent diode array detector at λ=254, 220 or 195 nm; room temperature.
8
NMR and HPLC Characterization Methods
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Example 1
General Methods:
1H NMR spectra were recorded on a Varian Inova 300 or 500 MHz NMR instrument. Chromatographic purities were determined on an Agilent 1200 Series or 1100 Series LC/MS system using a Merck Chromolith RP-18e analytical HPLC column (monolithic, 50×2 mm) and the following analytical HPLC method: injection volume 5 μL; flow rate 1 mL/min; 5→95% acetonitrile in water with 0.05% AcOH over 5 mins; Agilent diode array detector at λ=254, 220 or 195 nm; room temperature.
9
Spectroscopic and Colloidal Characterization of Nanostructures
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The 1H NMR measurements of TPEOH and SPOTPE in d6-DMSO were recorded on a 300 MHz spectrometer (Varian INOVA-300) at 25 °C. The chemical shifts were identified by the signals of tetramethylsilane (TMS). The ζ-Potentials of QC, SPOTPE, SPOTPE/QC and SPOTPE/QC+Fe3+ solutions were performed on a Nano-ZS ZEN3600 (Malvern Instruments, Worcestershire, UK) at 25 °C for three times to obtain the average values. The fluorescent spectra were measured on a spectrofluorophotometer (Perkin Elmer LS55) with an excitation wavelength at 336 nm and an emission wavelength at 472 nm. The excitation and emission slit widths were 10 nm and 4 nm, respectively. Photographs were obtained by a camera under a UV lamp with 365 nm excitation wavelength through the whole experiment.
Transmission electron microscopy (TEM) images were observed on a JEM-2100 (HR) electron microscope, using an accelerating voltage of 200 kV. TEM samples were prepared by dropping solutions onto copper grids coated with Formvar films, until the solvents were evaporated in a dust protected atmosphere. The dynamic laser light scattering (DLS) was performed on a light scattering goniometer (ALV/CGS-8F, ALV, Hessen, Germany) with a wavelength 632.8 nm from a He–Ne laser. The scattering angle (θ) was set at 90°. All of the sample solutions were filtered into the light-scattering bottles through the 0.45 μm filter (NYL, Whatman, UK).
Transmission electron microscopy (TEM) images were observed on a JEM-2100 (HR) electron microscope, using an accelerating voltage of 200 kV. TEM samples were prepared by dropping solutions onto copper grids coated with Formvar films, until the solvents were evaporated in a dust protected atmosphere. The dynamic laser light scattering (DLS) was performed on a light scattering goniometer (ALV/CGS-8F, ALV, Hessen, Germany) with a wavelength 632.8 nm from a He–Ne laser. The scattering angle (θ) was set at 90°. All of the sample solutions were filtered into the light-scattering bottles through the 0.45 μm filter (NYL, Whatman, UK).
10
In-vivo M. smegmatis Phosphate Analysis
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