We nurture a multidisciplinary research culture approach where new generations of researchers are versed in the most powerful techniques to understand biological problems at the level of the atom. A list of MSKC instruments and laboratory material available to registered users is provided below. For quick reference, users can find printed one-page guides located next to the instruments. MSKC staff provides training to first-time users.
Douglas Oryx8 Crystallization Robot
- Automated crystallization screening using nanoliter dispensing.
- Sitting-drop vapor-diffusion using Greiner CrystalQuick low profile, low birefringence, 96-well flat-bottom plates.
- Other sitting-drop plate settings available, e.g 1- or 2-samples per plate, with concave bottom Intelliplate.
- Further crystallization applications: microbatch under oil, LCP membrane protein crystallization. With/out seeding.
- 3-well crystallization experiments possible. Please consult first with the MSKC staff.
- Most popular commercial pre-made crystal screens, Silver Bullet and Morpheus additive screens available.
- Douglas Website.
Scorpion Liquid Handling Robot
- Art Robbins Instruments Scorpion liquid handler.
- Dispensing volume range: 1.0 µl to 1,000 µl.
- Design optimization screens based on initial crystal hits: pH-gradients, precipitant grids automatically calculated.
- Other liquid dispensing applications upon request, e.g. precise dispensing of precious aqueous solutions.
- Manufacturer Website.
GE Akta PURE FPLC
- Preparative purification system. Available columns:
- Ion-exchange: Mono Q 5/50 GL and Mono S 5/50 GL.
- Size-Exclusion: Superose 6 Increase, Superdex 75 and Superdex 200 Increase.
- Miscellaneous: HiPrep 26/10 Desalting, HiPrep Butyl FF 16/10 and HiPrep Phenyl FF 16/10.
- GE Healthcare Website.
Formulatrix RockImager2 UV-Vis Imager
- Dual visible light, UV light crystal plate imager, convenient tool to discriminate between protein and salt crystals.
- RockMaker software embedded for crystal hit scoring, link to crystallization screen conditions, X-ray diffraction data for scoring and tracking.
- Accepts all popular crystal plates: SBS-type, Linbro, Greiner, SwissSci.
- Stage holds up to two crystal trays, 5 minute scan per plate (visible light).
- Formulatrix Webpage.
Bio-Rad CFX96 Real-Time PCR
- Real-time protein melt analysis to explore chemical space in crystallography and cryo-EM.
- Thermal shift assay using Sypro Orange readout.
- Application: thermal denaturation screen for buffers that provide optimal two-state unfolding behavior.
- We provide the FUNDAMENT and SPECIFIC buffer screens (Jena Bioscience) which would help find stabilising the target protein/complex.
- Reference: Chari et al., 2015 Nature Methods paper: doi:10.1038/nmeth.3493
- Bio-Rad CFX Manager software available upon request.
MaxQ 8000 Orbital Shakers
Sorvall Lynx 4000 Centrifuge
- 6x 500 ml and 12x 50 ml capacity.
- Centrifuge max RCF 68,905 x g, max speed 24,000 rpm.
- Temperature range: -10° to +40°C
- Fiberlite™ F12-6x500 carbon fiber fixed-angle rotor (speed: max RCF 24,400 x g, max rpm: 12,000 rpm)
- Fiberlite™ F20-12x50 LEX fixed-angle rotor (max RCF 51,428 x g, max speed: 20,000 rpm)
- Thermo Fisher Website.
- 700W high-power sonicator, cell disruptor, homogeneizer.
- Processes 0.2 ml to 1 l.
- Ideal for cell disruption, homogenization, chip/DNA shearing, nano particle dispersion, protein extraction, sonochemistry and soil extraction.
- Fisher Website.
GE Amersham Imager 600
- Protein gel/membrane blot image acquisition instrument type 600 RGB.
- CCD 3.2 Mpixel camera fitted with a large aperture lens. Image resolution max.: 2816 × 2048, 5.8 Mpixel.
- Image output: Gray scale 16 bit tif, Color image jpg, Gray scale jpg
- Light sources: 460, 520, 630 nm (BGR), 470-635 nm (white), 312 nm (trans-UV)
- Chemiluminiscence, colorimetric epi-light, fluorescence trans-UV, epi-illumination, fluorescence red/green/blue epi light source.
- GE Healthcare Website.
Tissue Culture Essentials
- A variety of laboratory instruments complement of the TC room, including:
- A MaxQ 8000 shaker at 27℃ reserved for baculovirus culture (located in the corridor to the TC room).
- One Zeiss PrimoVert Microscope.
- One Thermo Sorvall ST16R centrifuge.
- Vortexer, water-heat bath, Neubauer cell counting chambers.
Rumed 3000 Crystal Incubator
- Rumed 3000 (Rubarth Apparate Gmbh) incubators set to one single-point temperature value at 6 °C and 16 °C.
- The heater and the fan are specially built to minimize variations in temperature, and its operation is essentially vibration-free providing an ideal environment for growing crystals.
- Why changing temperature as a crystallization parameter? Temperature influences nucleation and crystal growth by affecting the solubility and supersaturation of the sample. It has also been shown to be an important variable with phase separation in detergent solutions in membrane protein crystallization.
Discovery V12 Crystal Imaging Microscope
- Steroscopic microscope for crystal harvesting.
- Motorized 12:1 zoom controlled by the external touch panel SYCOP controller.
- PlanS 1.0x objective offering 8x to 100x magnification.
- SSRL SAM robot-compatible pins and loops for crystal harvesting. Other pins (e.g. ALS-compatible) available upon request.
- Zeiss Website.
Crystallography and cryo-EM Data Processing
- MSKC users have full control of the data they collect at SLAC. Users are trained to master tools to interpret results coming either from X-ray or electron beams.
- Current analysis software include CCP4 and Phenix. MSKC staff provides training.
Crystal Cryo-equipment and Crystal Harvesting Tools
- Complete selection of crystal harvesting tools including cryo pins, loops, casettes and all the necessary cryo-equipment.
- Standard cryo equipment includes CX-100 shipping dewar, VHC35 storage dewar, LD5 transfer dewar and standard SSRL equipment for use with the SSRL Automated Mounting (SAM) system using SSRL cassettes.
- MSKC is not a storeroom for crystals, it actively collects X-ray data thanks to rapid access to beamtime at SSRL/SLAC. Pleaase enquire for beamtime access through MSKC.
General Lab Equipment
- A variety of accessory and standard laboratory equipment are available including pH meter, heatblock, stirrers, rockers, rotators, microcentrifuges and microbalance.
- A heater/shaker block (Southwest Science) for either 50 ml conical tubes or 96-well deep well blocks or 96-well assay trays is available. Mixing speed: 200 to 1500 rpm in a 3 mm diameter vortex motion. Heating: ambient +5 C to 100 C.
- A high-speed, refrigerated microcentrifuge (Southwest Science): speed 200 to 15,000 rpm, up to 21,380 x g, is next to the Douglas for spin-down of protein.
- Thermo Scientific NanoDrop 2000c UV-Vis spectrophotometer. Quantify DNA, RNA, cell culture, and protein samples. Wavelength range: 190-840 nm. Minimum sample size: 0.5 µl. Cuvette mode for 1 ml cuvettes (0.5 ml minimum volume).
SDS-PAGE Gel Electrophoresis Mini Tank
- SDS-PAGE denaturing protein electrophoresis mini gel tank system.
- Runs with precast Bolt Bis-Tris Plus (Invitrogen) 4-12 % acrylamide gels: 8 cm x 8 cm (1.0 mm thick).
- Running buffer: MES SDS to resolve small-, medium-, to large-size proteins (0.3–260 kDa).
- In-gel detection on staining with PageBlue Protein Staining Solution (Fisher cat. # 24620). PageBlue is colloidal G-250 Coomassie dye, delivers a dynamic range of 5 to 500 ng.
- PageBlue is a safe, fast application dye (~ 1.5-2 hrs for SDS-PAGE gels), and it can be reused up to three times.
- Fisher Website.
Yeast Protein Production Platform
- UV-light sterilizable area for yeast culture, manipulation, inoculum/media preparation, plating.
- Equipped with automated pipettor head, anxillary tools.