DOE & Biochemistry Instrumentation Facility
The UCLA-DOE Instrumentation Core and Biochemistry Department Shared Instrumentation facility work to provide the UCLA research community open access to sophisticated equipment for biochemical studies. These facilities house a wide array of detection systems useful for analysis, quantification, and identification of all types of macromolecules and their molecular interactions. It also carries equipment for cell culturing, harvesting, lysis, and high-speed centrifugation useful for biomolecule preparation. Follow the links below for more information and to contact facility managers for training.
Multi-Angle Light Scattering (MALS/T-rEX)
The Wyatt miniDawn TREOS is a multi-angle light scattering detector (MALS), reading at three different angles. This instrument is capable of calculating molecular weights and average dimensions of macromolecules and complexes in solution with great accuracy. The system works in tandem with an Optilab refractometer with extended range (T-rEX), to determine the absolute concentrations of analytes. When coupled with gel-filtration chromatography, this MALS/T-rEx combination system can easily detect the molecular weight, spatial size, and concentration of macromolecules in mixed solutions. This allows users to identify, characterize, and determine relative abundance of complexes formed by mixed analytes, or the oligomerization states of single proteins. As currently operated, our system is suited to aqueous systems and relatively large molecules only.
Isothermal Titration Calorimeter (iTC-200)
The Microcal iTC-200 is designed to make very precise calculations of the binding energetics of analyte molecules, most commonly a protein and ligand. Through precise measurements of enthalpy changes produced by titration of one analyte into the other, this instrument can calculate binding constants and stoichiometry with a high degree of accuracy. This machine uses a far smaller sample volume than previous models (only 200 L), opening this instrument to experiments with analytes that are difficult to produce in quantity.
Pharos FX Plus Molecular Imager
The Pharos FX system is a robust yet simple system for visualizing and quantifying a wide array of gels, blots, and screens. This Pharos FX is capable of visualizing samples by fluorescence, visible light/colorimetric, and phosphorescence (radiography screens). It is also capable of multiplexed reading, expanding the complexity of available experiments. In addition to the standard internal 532nm laser, our system incorporates external lasers at 488m and 635m, further increasing applicability of this work-horse system to the full range of fluorescent and colorimetric dye sets. The facility also has an older, though equivalent, BioRad FX ProPlus scanner rated for use with radioactivity and best suited for scanning of phosphorescent screens, though most applications are possible with this system.
Tecan M1000 Plate-Reader
The Tecan Infinite M1000 Pro is a multi-wavelength automated plate-reader system. It is ideal for many types of light based high-throughput assays: from simple growth-curves to more complex luminescence and fluorescence experiments, both in single read and kinetic formats. The system is equipped with an automated injector for uninterrupted kinetic assays, as well as Top and Bottom read modes to accommodate solution or cell and surface based assays. The Tecan Quad4 Monochromator system finely tunes excitation and emission light to any wavelength without standard filters, from near-infrared to Ultraviolet. This light system sacrifices a small degree of signal intensity to allow users complete control of the assay wavelengths as well as the bandwidth of the light produced and read.
All data is automatically exported into a Microsoft Excel spreadsheet, allowing for simple data processing without the requirement of specialty Tecan software.
Spectrofluorimeter (PTI QuantaMaster)
The Photon Technologies International QuantaMaster Spectrofluorimeter allows for a large range of both single-read and kinetic light-based assays. This systems is capable of producing and measuring through the entire range of wavelengths from 260-700 nm. Additionally, onboard polarizers allow for fluorescence anisotropy experiments to be performed. Beyond single frequency reads, the QuantaMaster is also able to scan through a desired range of emission and excitation wavelengths. Our facility provides users with a limited supply of various sized quartz cuvettes for experimentation, including small volume cuvettes for difficult to produce or expensive analytes (some samples may require users to provide their own cuvettes).
Bio-Rad CFX Connect: quantitative PCR
The CFX Connect qPCR system enables real-time detection of DNA copies amplified during PCR through the use of intercalating or primer based fluorophores.
Quantitative PCR provides sensitive measurements of gene transcription levels, detects gene mutations down to the single base level, among many more nucleic acid probe based experiments; it can also be applied to protein stability assays using the Sypro-Orange dye. The samples are measured with a trio of excitation LEDs and filtered emission detectors covering the range of 450 to 580m; comprising nearly all commonly used fluorescent dyes and probes, with a dynamic range of 10 orders of magnitude.
BioXp 3200 Gene Synthesizer
The BioXp 3200 is an automated platform that rapidly synthesizes high quality linear DNA fragments from custom designed oligo-pools across a large complexity continuum. In a single run, this system can produce up to 31 fragments ranging from 400 bp to 1.8 kb in length. DNA fragments are ready for sequencing QC, further assembly into larger fragments, cloning into vectors and other common downstream applications. Future modules will allow for direct cloning of fragments into vectors, integrated onboard the BioXP within a single run.
The UCLA-DOE & Biochemistry Instrumentation Facility understands that groups interested in using this platform may need only a small number of DNA fragments synthesized, and thus are reluctant to use this powerful tool. In order to facilitate our mission to the UCLA research community, we are offering to pool small orders into single plates. This will help share costs of reagents and consumables among labs, and allow groups access to the exceptional rate of $0.12 per base pair being offered to us by Synthetic Genomics, Inc.
For information and instructions on having your
sequences synthesized with the group-ordering platform, see below:
BioXp 3200 Gene Synthesizer – Pooled Ordering
SGI-DNA is not currently charging for shipping, pipet-tips and other consumables necessary for using the BioXP. Therefore, pooled ordering is unnecessary as there are no savings for users by grouping orders; you are essentially only paying for your DNA. If in the future such items are no longer complimentary, the facility will institute pooled ordering once again.
LiCOR Odyssey Infrared Scanner
The LiCOR Odyssey is a highly sensitive scanner operating in the infrared spectrum. This system is primarily used for western blot analysis, though its applications include EMSA/gel shifts, DNA-stained gels, and more. While this requires the use of specialized antibodies and detection products, the benefits of the system far outweighs the cost. Direct imaging of conjugated antibodies eliminates film costs and allows for a wider linear dynamic range of quantification than chemiluminescence. The LiCOR also achieves a higher signal to noise ratio than other detection methods, due to negligible background radiation from the membrane at the signal wavelengths. For peak performance, users are suggested to use membranes made of nitrocellulose or Millipore Immobilin-FL membrane a low fluorescence PVDF.
Stopped-Flow Spectrometer
Our Applied Photophysics SpectroKinetic Monochromoter stopped-flow spectrometer measures fast molecular reactions over a wide-range of signal wavelengths. Sample mixing occurs within 2 milliseconds, with large pools of signal data recorded on the order of 10 milliseconds; longer time courses are also possible on this instrument. The system is equipped with temperature controllers when needed.
Circular Dichroism Spectrophotometer
Our JASCO 1-715 circular dichroism spectrophotometer is a robust instrument used to determine the secondary structural elements of macromolecules, as well as their stability and conformational changes under a wide range of conditions. Our JASCO CD spectrophotometer is also equipped with a Peltier temperature controller and an auto-titatration module that greatly expands the range of experiments which may be performed.
Differential Scanning Calorimeter
Our facility houses an Applied Themodynamics differential scanning calorimeter (Model: N-DSC II). This instrument is used to study the phase-transititions and stability of macromolecules over a range of temperatures, additionally, it can be used to study the interactions of a macromolecule with ligand binding partners. The technique of DSC compares the heat required to make defined temperature changes between a sample chamber and reference cell, and thus is able to determine a wide-range of thermodynamic properties of the tested system.
Agilent BioAnalyzer 2100
The BioAnalyzer is an electrophoresis-based analytical system capable of rapid sizing, quantification, and quality control of biomolecules. Our facility provides probes for DNA and protein samples, as well as a dedicated probe for the analysis of RNA; we do not currently support the flow-cytometry capabilities of the BioAnalyzer system. Sample chips and reagents are not provided by the facility.
Bio-layer Interferometry (BLItz)
Bio-layer interferometry uses the interference produced from two light reflections of a single source to measure the aggregation of a target molecule on the sensor surface: as the target molecules aggregate or dissociate from the probe surface, the distance of between the reflections sources change accordingly. Simple dip and read measurements can be used to identify the presence of a molecule in a solution, while time-course analysis allows for measurements of binding affinities. A wide range of probes are available (standard antibodies, Protein A, Ni-NTA, Streptavidin, etc.) making this system extremely flexible in application; furthermore, limited custom probe production is possible in coordination with the manufacturer.
Hewlett-Packard UV-Vis Spectrometer
The HP UV-Vis is a standard spectrophotometer capable of single-read and kinetic measurements. The system is also equipped with an onboard Peltier Temperature Controller that allows for samples to be maintained at single temperatures, or read through series of temperatures (such as nucleic acid melting experiments). A basic set of cuvettes are provided for users, though certain samples may require researchers to provide their own.
Nano-Drop Spectrophotometer
The Nano-Drop by is a fast-read out UV-Vis Spectrophotometer suitable for analysis of relatively high-concentration samples, requiring only a small sample of 1-2uL. This instrument is ideal for the fast determination of the concentration or purity of nucleic acids and protein samples prior to other experimentation.
Alphalmager FC2 CCD Camera
This versatile CCD camera system produces high-quality images of a wide range of samples. It is equipped with both visible and ultraviolet epi and transmission light allowing for the imaging of nucleic acid gels and visibly stained protein gels. The visible light sources on this camera also allow for the imaging of bacterial and cell culturing plates. Finally, this system is equipped with luminesce capability allowing for the imaging of light producing samples, including but not limited too, westerns using chemiluminescent secondary antibody systems.
Nexcellom Auto M10 Cell Counter
This simple and rapid cell counter is ideal for the quantification of eukaryotic cells in a wide range of applications. Additionally, the instrument requires only a small sample size, and produces both high-quality images as well as multiple analysis and quantification modes. Special slides are required, and may be purchased through the instrument manufacturer.
High-Speed Centrifugation
Model: Avanti J-25
Top Speed: 25,000 rpm
Cooling: Yes
Rotors Provided: JA-10, JA-14, JA-25.50
Ultra-Speed Centrifugation
Model: Optimax Max-E
Top Speed: 100,000 rpm
Cooling: Yes
Rotors Provided: TLA-110 (one rotor)
Model: Optimax TLX
Top Speed: 120,000 rpm
Cooling: Yes
Rotors Provided: TLA-110 (one rotor)
Model: Beckman LE-80
Top Speed: 55,000 rpm
Cooling: Yes
Rotors Provided:
Microbial Culturing
Our facility is equipped with two large incubator shakers, ideal for 500mL or greater liquid cultures, and two smaller bench-top models ideal for tubes and smaller cultures. All units are equipped with variable speed and temperature controls. We also house a large box incubator for culture plates or other solid surface culturing.
Cell Lysis
Our facility houses a Sonicator system equipped with fully programmable pulse and timer controls, including exchangeable tips to accommodate individual experimental needs. We also have a high-pressure Microfluidizer capable of achieving 90% cell lysis in two-passes through the system. Finally, we house a traditional French Press system.
Training
TRAINING BY FACILITY SUPERVISORS IS REQUIRED BEFORE USING ANY and ALL INSTRUMENTS
Instrument trainings are arranged with one of the Instrumentation Managers. Trainings will take 30 minutes to 2 hours for more complicated instruments. Topics will include the basic principles and applications of the instrument, proper handling and usage, and data production and analysis when necessary.
If you wish to receive training on instruments in our facility, contact the Instrumentation TA (use link above), and provide the following information:
- Your Name and research level (Undergrad/Grad/Postdoc)
- Research Supervisor’s Name / Lab Group
- Name of Instrument(s)
- Basic information about desired use or experiment, if possible.
Contact
Biochemistry Shared Instrumentation Manager
Matthew A.G. Graf
bita@chem.ucla.edu
UCLA-DOE Instrumentation Core Manager
Martin Phillips
mlphill@chem.ucla.edu
Faculty Instrumentation Committee Chair
Do Not Contact for Trainings or General Information
Margot Quinlan
margot@chem.ucla.edu
Reservations
All Users must be trained by facility management prior to making
reservations on any instrument.