Cell and Molecular Biology
Objectives
The Cell and Molecular Biology Core:
- Provides access to relevant technologies and equipment
- Provides education, training and consultative advice regarding these technologies and equipment
- Provides financial subsidies for key technologies
Specific Services
Advice and Training in Cell and Molecular Biology Techniques
The core can provide advice regarding the design and interpretation of experiments
using basic molecular techniques such as PCR, transfections, etc. The core
can assist with the computer analysis of DNA or protein sequences. Users
who have highly specialized needs that require expertise beyond the core
will be referred to other appropriate laboratories where feasible.
Transgenic Animal and Vector Core Facility Subsidies
The MDRTC Cell and Molecular Biology Core can provide limited subsidies
(typically 25% of cost, though the total subsidy for each laboratory may
be capped) to users of the Biomedical
Research Council Transgenic Animal and Vector
Core Facilities. A Transgenic Animal Core Subsidy
Application or a Vector Core Subsidy Application
should be completed and submitted via the Web. The application will
be reviewed, and you will be notified of approval/ disapproval. To be potentially
eligible for the subsidy, the project must be relevant to diabetes
or related endocrine disorders, and this relationship must be clearly
justified on the application form. For the purpose of these applications,
“related endocrine disorders” are those that impact diabetes,
such as obesity.
Microarray Analysis
Gene expression profiling using Affymetrix GeneChips is available at a discounted
rate through the Cancer Center
Microarray Facility. To be potentially eligible for the subsidy, a Microarray
application form should be filled out and submitted via the Web. The
application will be reviewed, and you will be notified of approval/ disapproval.The
project must be relevant to diabetes or related endocrine disorders,
and this relationship must be clearly justified on the application form.
For the purpose of this application, “related endocrine disorders”
are those that impact diabetes, such as obesity. Microarray analysis is
very expensive. The MDRTC typically will provide a 25% discount, though
the total subsidy for each laboratory may be capped.
Hybridoma Core
The MDRTC will continue to provide subsidies for use of the Hybridoma Core,
typically equal to 50% of charges, though the total subsidy for each laboratory
may be capped. Please see the Hybridoma Core web site. To be potentially
eligible for the subsidy, the project must be relevant to diabetes or related
endocrine disorders. “Related endocrine disorders” are those
that impact diabetes, such as obesity.
RNA interference / shRNA lentiviral plasmids
The UM Functional Genomics Core (3338 LSI; 615-5309; http://fgc.lsi.umich.edu)
houses, maintains and distributes lentivrial plasmid vectors expressing
shRNAs directed against most mouse and human genes, to be used in RNA interference
gene silencing studies. The MDRTC Cell & Molecular Biology Core will
provide a 50% subsidy toward the purchase of these shRNAs, though the total
subsidy for each laboratory may be capped. An application form is not necessary.
There should be an MDRTC check box on the UM Functional Genomics Core request
form, which will trigger the subsidy.
High Throughput Chemical Screens (Center for Chemical Genomics,
3338 LSI, 615-9422, http://www.lsi.umich.edu/ccg)
The Center for Chemical Genomics performs high throughput robotics screens
of a library of ~50,000 small molecules to identify compounds that activate
or inhibit a biological process of interest. Typical goals are to either:
a) identify novel chemicals to rapidly activate or inhibit the process and
study the biological consequences; or b) find potential leads for the identification
of useful drugs. The MDRTC can subsidize 1 or 2 users per year at 50% of
the cost up to $10,000 per PI. (We will subsidize the actual small molecule
screen, after assay development is complete). The project should be discussed
with the CGC directly to assess feasibility and to develop the screening
assay. The CCG form contains an “Other Funding Information”
box; you apply for the subsidy by writing MDRTC in that box. To be potentially
eligible for the subsidy, the project must be relevant to diabetes or related
endocrine disorders. “Related endocrine disorders” are those
that impact diabetes, such as obesity.
Bacterial Artificial Chromosome Recombineering Core
Bacterial Artificial Chromosomes (BACs) are plasmid-like constructs
that can be grown in E. coli and that contain genomic DNA fragments of ~200
kb. BACs are commercially available that contain most of the genes from
human, mouse and other species. These can be modified (“recombineered”)
by insertion, deletion or point mutation, and can then be used as targeting
vectors to modify endogenous genes in mice, or to produce transgenic mice.
Thus, BACs can be used to create mouse models of human diseases and to make
mice expressing reporter genes. Since BAC recombineering is technically
demanding, the BAC Recombineering Core has been established.
The BAC Recombineering Core is fully operational as of October 2009.
This core is managed by Thom Saunders tsaunder@umich.edu
and functionally is part of the Transgenic Animal Model Core. The MDRTC
provides substantial support for the BAC Recombineering Core, therefore
MDRTC members utilize its services at a significant discount. MDRTC investigators
will be charged $500 for BAC recombineering (per construct), and $25 for
BAC DNA purification including pulsed field gel verification of restriction
enzyme fragments. The BAC Recombineering application should be completed
and sent to 2560 MSRB II, Box 0674.
Equipment
The equipment described below is available to MDRTC members and is located in room 5570-E MSRB-II. Room keys cannot be provided to users, and the room must be kept locked when it is not in use. We regret the inconvenience, but this means that users may not have reliable access during non-business hours. Users must first receive training by Dr. Ron Koenig before they will be able to access the equipment.
- BioRad Phosphorimager. The Phosphorimager creates a digital image of radioactive gels, blots, etc. that is amenable to quantitative analysis and the production of electronic slides or figures. All common isotopes except tritium are detected. After receiving training, labs will be given a password that is required for instrument operation.
- BioRad Fluor-S Max Imager. The imager is essentially a phosphorimager for chemiluminescence, such as Western blots, using ECL technology. After receiving training, labs will be given a password that is required for instrument operation.
- Applied Biosystems StepOnePlus real time PCR machine. After receiving training, users can sign up for this machine. Sign-up rules and instructions are near each machine.
The following equipment is available to MDRTC members in room 6235 of the Brehm Tower:
- Homodyne Luminary/FX Chemiluminescence Western Blot Detection System. The Western Blot Detection System allows MDRTC members to image and accurately quantify (over many orders of magnitude), various types of blots, using luminescent/fluorescent/chemiluminescent technology.
- Scintillation Counter. The scintillation counter is available for use in radiometric assays for enzyme activity, binding, etc.
- Applied Biosystems 7500 real time PCR machine. After receiving training, users can sign up for this machine. Sign-up rules and instructions are near the machine.
Key Personnel
Ronald J. Koenig, M.D., Ph.D., Core Director, rkoenig@umich.edu; 734-615-9497


