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FACILITY INFORMATION
Facility Director
Anne V. Kane, M.D.
akane@tufts-nemc.org
Technical Staff
Lynn Gorczyca
Location
Tupper Building, Room 822 15 Kneeland Street
Tufts-New England Medical
Center
Mailing Address
750 Washington Street, Tufts-NEMC
Boston, MA 02111
Telephone
617-636-7611 |
We provide services and reagents, at competitive prices,
for molecular biology and microbiology research. Our services
are available to all Tufts scientists.
Large-scale (up to 20L) Culture
The Intestinal Microbiology Core has large-scale bacterial culture and processing
capability using the NewBrunswick Bioflow 4500 fermentor. 20L cultures of aerobes,
anaerobes, or microaerophilic bacteria are readily produced. Our cell press,
originally purchased for lysis of large-scale culture pellets to release cytoplasmic
proteins, can also be used to process small batches when sonication
is an undesirable option. A Pellicon tangential flow filter apparatus is used
for rapid harvest of bacteria from large culture volumes, and in parallel with
ultrafiltration membranes, to reduce culture volumes. This rapidly concentrates
culture supernatants from 20L to 500ml, facilitating purification of secreted
proteins. The cell press and the Pellicon apparatus are available
to scientists for either small- or large-scale cultures.
Ordering
Scientists who wish to order large-scale cultures should contact the core
laboratory facility director to discuss scheduling. Cultures>10L require
prior approval from the Institutional Biosafety Review Committee. We try to
deliver within two weeks following receipt of the strain. The culture can be delivered either as a cell pellet or as the supernatant
following ultracentrifugation of the cell lysate. If
the desired product is in the culture supernatant, we can reduce the volume
to 500ml and wash with any buffer desired. We require a plate with the organism
of interest, IPTG if required for induction, and a protocol that should include:
- growth
conditions and medium to be used
- information
regarding antibiotic resistance
- desired
culture optical density prior to induction
- duration
of incubation post induction
- buffer
for wash, if desired
- preferred
lysis buffer, and protease inhibitors, if desired
Culture of Pathogens
The core laboratory in Tupper 822 contains all the instrumentation required
for culture, including an autoclave, incubators, water bath shakers, and the
20L fermentor. The facility is certified at a P2/ large culture level of containment,
and appropriate lab practices are strictly observed.
Organisms
provided over the past five years include enterohemorrhagic E.
coli, Vibrio cholera, Yersinia pseudotuberculosis, Legionella
pneumophila, Clostridium difficile, Hemophilus
influenzae, and Helicobacter pylori. The latter
organism requires special chambers for the microaerophilic
environment, and 10 such chambers are available in the
core. In addition, the core has the equipment and expertise
to culture Giardia lamblia and Entamoeba histolytica in
large culture volumes. Stocks of both these parasites are
maintained in liquid nitrogen. Requests for culture should
be made to the core laboratory facility director; the price
is based on the cost of the medium used.
Storage and Retrieval
The core maintains a large collection of bacteria (frozen stocks) available
as a resource for all investigators. This library is on a database on the core
computer and can be sorted using a number of identification properties. The
basic collection includes pathogenic bacteria, several standard host
strains, and commonly used plasmid vectors. The core
also maintains microorganisms for --these are the sole repository
of bacteria for some laboratories, or are a second storage site to prevent
loss of valuable strains.
Requests
for bacteria to be delivered on plates can be made by phone
or e-mail, and delivery is usually on the following day.
For bacterial or plasmid storage we require a plate streaked
for isolation. There is no charge for this service.
Services
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