Services Provided by the MRCF
The MRCF offers a wide variety of molecular services. We accept whole tissues all the way through library prepped samples, and are willing to assist at any step of the way for preparing your samples for analysis. We are always willing to help, and if you are looking for a service that you don't see listed, let us know and we will discuss the option of optimizing and adding it to our list of services we can provide.
If you have any additional questions, please call us at (208) 282-4148 or email our lab email, email@example.com.
Next-Generation DNA Sequencing
Next-Generation Sequencing (NGS) brings a new tool to science that is opening doors in all scientific fields. Just in microbial genomics, NGS allows for characterizing of un-culturable microorganisms, discovery of new viruses, and provides insight into new outbreak control methods.
Here is a list of just a few of the applications NGS is used for:
Complex Disease Genomics
Microbial Genomics and Environmental Profiling
Genomics in Drug Development
Reproductive and Genetic Health
NGS has revolutionized how genomes and DNA sequencing can further scientific research. Read more at Illumina's website for project design ideas and inspiration.
The MRCF provides library preparation of all types for NGS on Illumina instruments, as well as sequencing on any instrument.
The cost varies greatly based on the project design. Please contact the MRCF for a quote on your particular project.
Sanger DNA Sequencing
Sanger sequencing is performed on our AB 3130xl 16-capillary genetic analyzer. We can process 1-96 samples per plate of samples submitted, and will include one pGEM control sample either per run, or per plate.
Please submit a Job Request Form, see link below, with your samples. This is a downloadable Excel file that you will fill out and email an electronic copy to firstname.lastname@example.org, as well as a send a hard copy along with the samples submitted to us.
First time customers, will need to submit a one-time New User Form.
Internal Rate: $4.00 / sample; External Rate: $5.00 / sample
We can quantify and dilute the samples to the appropriate concentrations if needed. For an additional $1.00 / sample, we can cleanup PCR products.
Each sample submitted needs to have 5uL of template DNA diluted to the appropriate concentration for the DNA sample type. Add 1uL of 3.2uM primer (only one) in each sample for a total volume of 6uL to be submitted to the MRCF. Visit our page for more details on how much sample to use and what concentrations are needed for different types of DNA sequenced.
Fragment Analysis - Microsatellites and DNA/RNA Profiling
3130xl Fragment Analysis - Microsatellites and T-RFLPs
Microsatellite markers, also known as short tandem repeats (STRs), are polymorphic DNA loci consisting of a repeated nucleotide sequence. The number of repeats varies in a population, thus creating multiple alleles for a microsatellite locus. Microsatellite loci are amplified by PCR using fluorescently labeled forward primer. These PCR amplicons are then separated by size on the AB 3130xl genetic analyzer. T-RFLP analysis is a technique used to study complex microbial communities based on variation in the 16S rRNA gene. It can be used to study microbial community structure, bacterial populations in the environment and changes in a microbial community in response to different environmental parameters.
Our instrument runs batches of 16 samples - but entire plates can be set-up and scheduled to run on the instrument.
Please submit a Job Request Form, see link below, with your samples and specify the size standard you want us to use. This is a downloadable Excel file that you will fill out and submit electronically to the MRCF, as well as send a hard copy along with the samples submitted to us.
First time customers, will need to submit a one-time New User Form.
Available Size Standards: ROX350, LIZ600, ROX1000
Please send a minimum of 5ml of sample. For a new fragment analysis project, we recommend that several stock dilutions be run on the sequencer to determine the optimal sample dilution, for example 1:10, 1:100, 1:200.
$2.00 / sample
DNA/RNA Profiling by Advance Analytical's Fragment Analyzer
The Fragment Analyzer covers the full spectrum of DNA, gDNA and RNA quality control. It is used and recommended by Illumina as an automated system for the quantification and qualification of NGS libraries, gDNA and RNA. Our instrument has 12-capillaries that electrophoreses 11-fluorescently labeled samples and a size standard (ladder) per instrument run. With a run time of a little less than 1 hour, it resolves fragments from 10-40,000 bp. The instrument has a resolution of 2bp for fragments which can be detected in as little concentration of 5 pg/uL. The instrument is one of the best to look at the profile, integrity and quality of samples.
$2.50 / sample (internal and external)
Microscopy and Histology
Multiphoton (2P) Microscope, Olympus FV1000MPE
The multiphoton microscope excitation is superior for imaging living cells that reside within intact tissues such as brain slices, embryos, whole organs, and even entire animals. The technique provides optical sectioning with reduced absorption of excitation light in specimen regions removed from the objective focal plane, thus minimizing photobleaching and phototoxicity. Please talk to MRCF staff to see if this system is right for you!
Confocal Microscope, Olympus FV1000
Our confocal laser scanning microscope has four simultaneous detection channels, including one for DIC and two spectral channels. The lasers used for excitation on our instrument cover the visible spectrum; the exact wavelengths are 405nm, 458nm, 488nm, 515nm, 559nm and 635nm. It is important to talk to one of our staff members before setting up an experiment to make sure there won't be crosstalk or crossing of two spectrally-similar colors.
Brightfield Microscopy - Upright Leica DM6B
The DM6B is a fully automated microscope capable of brightfiled, darkfield, DIC, phase contrast, and epifluorescence imaging. This scope can also simultaneously capture fluorescence and DIC, and fluorescence and phase images. The ease of use makes this microscope ideal for teaching courses, and great for quick sample screening.
Transmission Electron Microscopy - Zeiss EM900
The Transmission Electron Microscope (TEM) is capable of imaging ultra-thin specimen at the magnification of 400,000X, - easily detecting targets as small as cellular organelles and viruses. Use of the TEM requires meticulous sample preparation of tissue support mesh grids coated in a layer of carbon molecules. Our newly acquired Gatan CCD digital camera makes image capture quick and easy. Sample preparation is specialized for this instrument, please contact MRCF staff with questions about this process.
Histology Equipment and Offline Data Analysis Station
Cryostat, Leica 3050 S
When working with delicate specimens the precise specimen orientation and the specimen feed system of the Leica cryostat guarantees reproducible, thin, serial sections of maximum quality. The 3050 S is the top of the line model for cryostats, and is capable of automatic, foot-pedal driven sectioning for convenience of the user. A second cryostat, the Leica 3050, is also available for use.
Microtome, Leica RM2255
Designed for fully motorized sectioning of both paraffin- and resin-embedded specimens, the Leica RM2255 supports a broad spectrum of applications.
Offline Data Analysis Station
This offline workstation includes multiple programs to help researchers analyze data generated from the Advanced Imaging Facility, and produce publication worthy images and pictures for various uses. Programs available on this computer are MetaMorph for Olympus, ImageJ (Fiji), Vaa3D - 3D Visualization-Assisted Analysis, Volocity Demo 6.1.1 and FluoRender. In addition to analysis software, the computer includes Microsoft Office and Adobe Reader.
Flow Cytometry is the analysis of fluorescent markers and dyes on virtually any cell or cell-sized particle. It is a simple, rapid and powerful tool that takes advantage of fluidics, laser and PMT technology, and high tech electronics for detection and analysis of fluorescence.
The FACSCalibur incorporates efficient optics and two air-cooled solid-state lasers to analyze up to four colors. Compact and easy to use, the FACSCalibur is a highly sensitive tool, providing researchers with the technological capabilities crucial for sophisticated cellular and molecular work in areas such as cell cycle, cell function, multicolor analysis, and immune function.
Our instrument has a mechanical sorter that allows researchers to sort out cell populations of interest, and save them for other applications. The sorter is able to sort at around 2,500 cells/second, which is fast enough to collect large numbers of cells in a fairly short time, and keep them viable and ready to use.
Currently, there is no charge for using the Flow Cytometer. We have excess reagents and a fully funded service contract. We encourage our researchers to take advantage of this while it lasts!
Read this brief Flow Cytomety Tech Note on how to use flow to quantify the amount of living cells versus dead cells in a sample.
The MRCF is provides a wide diversity of applications, all at one core facility! If there is a service you require, that isn't listed below, please contact us - we are more than happy to develop and optimize new techniques.
ELISA Immunosorbent Assays
qPCR and PCR