Equipment Core

Equipment is available to Department of Pediatrics investigators and their laboratory personnel. 

Location: Room 8240 MSRB III 
First two benches through the door across from the elevators.

Hours of Operation:  Monday - Friday, 8:00 AM - 6:00 PM
Contact: Linda Kalikin, PhD - [email protected]
Scheduling: Reservation calendars are posted next to qPCR machines. All other equipment is available on 'first-come / first-serve' basis.
Access to computers controlling individual instruments is restricted. Please email the Core Director ([email protected]) to be added to the Facility’s “user list”.

 


 

FluorChem M MultiFluor System

The FluorChem M MultiFluor System (Protein Simple) performs quantitative multiplex fluorescent and chemiluminescent imaging of Western blots as well as protein stained gels. Instrument runs on AlphaView software ver 4.1.4 which offers tools for capturing, analyzing, and annotating images. Stand-alone software is available for download to PC computers.
User Manual:  FluorChem M manual.pdf 
Software Guide:  AlphaView User Guide_for FluorChem.pdf

** No imaging of ethidium bromide stained gels on this instrument. Please use the BioRad ChemiDoc imaging system. **

  

The StepOnePlus Real-Time PCR System (Applied Biosystems) is a 96-well PCR instrument capable of endpoint and quantitative assays. Instrument accommodates both TaqMan and SYBR Green reagents and includes four filters calibrated for FAM/SYBR Green, JOE/VIC, TAMRA/NED, and ROX dyes allowing for the design of a variety of singleplex assays as well as multiplex assays with up to four colors. VeriFlex block enables up to six different temperature zones. StepOne Software ver 2.3 supports analyses by standard curve /absolute quantitation, relative standard curve, comparative Ct (DDCt) /relative quantitation, presence-absence, and melt curve. User Manual:  ABI StepOnePlus Manual.pdf  
 
Eppendorf qPCR_2

The Mastercycler ep realplex2 Gradient Thermocycler (Eppendorf) is a real-time PCR system with high ramping speeds and short detection times. LEDs generate a blue light at a wavelength of ~470 nm, which can excite nearly all fluorophores used in real-time PCR, including SYBR Green, FAM, VIC, TET, HEX, ROX, JOE, TAMRA. Instrument incorporates two fluorescence filters and one photo-multiplier tube (PMT) allowing duplex assays to be performed.
User Manual Epp Mastercycler ep realplex user manual.pdf 
Software Guide:  Epp Mastercycler ep realplex software guide.pdf   

 

 

 

  The Gel Doc XR+ Imaging System (BioRad Laboratories) enables quick and easy visualization, documentation, and analysis of nucleic acid and protein gels, blots, and macroarrays using high-resolution, high-sensitivity CCD detection technology. System supports a range of fluorescence and colorimetric detection methods, including ethidium bromide; SYBRs Green, Safe, and Gold; Coomassie Blue; Flamingo; Oriole; Krypton; copper, zinc, and silver stains; and Qdots 525, 565, and 625. Image Lab Software (ver 6.0.1) annotates and documents images, analyzes molecular weights and base pairs, and determines accurate quantitation and purity of samples. Images and data can be printed using the attached thermal printer or exported for use with other software such as Microsoft Office programs.  User Manual and Software Guide  

 

 

SpectraMax M2e Microplate Reader

 The SpectraMax M2e Microplate Reader (Molecular Devices) combines the UV-vis detection of a spectrophotometer and the fluorescence detection of a spectrofluorometer in one instrument. Instrument accommodates cuvettes and 6, 12, 24, 48, 96, or 384 -well plates. Absorbance range is 200-1000 nm, tunable in 1 nm increments. Fluorescence detection utilizes dual monochromators, top or bottom read, wavelength scanning, and reads up to 4 wavelength pairs in one protocol for endpoint and kinetic measurements. Secondary modes available for limited development of glow luminescence or time resolved fluorescence assays. Applications include DNA / RNA / protein quantitation, ELISAs, enzyme kinetics, FRET with fluorescent proteins, cell viability & cytotoxicity, reporter genes, cell migration & adhesion, fatty acid uptake, membrane permeability, and microbial growth assays. Reader runs on MD’s SoftMax Pro Data Acquisition & Analysis Software (ver 5.4).  
User Manual:  spectramax_m2_Mol Dev User Manual.pdf

 

PHERAstar FS Microplate Reader

The PHERAstar FS Microplate Reader (BMG Labtech) is a multi-mode microplate reader, however this unit is configured only for use with PerkinElmer’s Amplified Luminescent Proximity Homogeneous Assay (Alpha) Technology, a bead-based proximity system that measures biological interactions with more sensitivity and less time than other methods (e.g. ELISAs). Alpha Technology is suitable for biochemical and cell-based assays, low to high affinity binding interactions (pM to mM), high-throughput screening (HTS), and complex matrices such as cell lysates, serum, plasma, and CSF. AlphaScreen assays provide convenient off-the-shelf solutions for fusion tag detection, while AlphaLISA assays offer a wide range of beads for flexible mix and match. Applications include protein-protein and protein-nucleic acid interactions, ligand-receptor, kinase, epigenetics & post-translational modifications, protease, apoptosis, and second messenger cAMP/cGMP assays. Instrument runs on MARS Data Analysis Software ver 2.40. 
User Manual PHERAstar FS Operating Manual.pdf
Software Guide PHERAstar Software Manual.pdf
Data Analysis PHERAstar_data analysis.pdf

 

Stratalinker 1800

The Stratalinker® UV Crosslinker (Strategene) is designed to crosslink DNA or RNA to nylon, nitrocellulose, or nylon-reinforced nitrocellulose membranes for Southern, northern, dot, or slot blot analysis and colony or plaque screening. Other applications include nicking of DNA in agarose gels prior to blotting, dimer formation to perform partial digests for rapid restriction mapping, UV sensitivity testing for host strain verification, and irradiation of PCR reagents, pipets, and tubes to prevent introduction of exogenous DNA into reactions.
User Manual:  Stratagene Stratalinker manual.pdf  

 

Bio-Rad/BioRad SmartSpec/Smart Spec 3000 UV/Visible Spectrophotometer

 The SmartSpecTM 3000 Spectrophotometer (BioRad Laboratories) is a scanning tabletop UV/visible spectrophotometer that measures absorbance from 200 nm to 800 nm. Applications include quantitation of DNA, RNA, oligonucleotides, and proteins, monitoring of bacterial culture growth, simple kinetic assays, and wavelength scans with peak detection. Spec is capable of performing calculations and modifying/storing conversion factors to generate A260/A280 ratios for nucleic acid purity, sample concentrations that take into account dilutions factors, and molar extinction coefficients. Instrument also features a built-in printer.
User Manual:  BioRad spectrophotometer manual.pdf 

Locations

8240 MSRB III