Susann Brady-Kalnay, Ph.D.,
Module Director
Department of Molecular Biology & Microbiology
phone #368-0330
Moonkyung Hur Caprara, Ph.D., Module Manager
Department of Ophthalmology
phone #368-4990
Located in The VSRC Consolidated Laboratory
Space in the Institute of Pathology
The Molecular Biology (MB) Module will facilitate
the use of fundamental techniques in molecular
biology for the many vision investigators who
desire to add these tools to their research
capabilities as well as for investigators who
already use some of these techniques . The
MB Module is responsible for providing skilled
technical expertise, equipment and techniques
for PCR analysis/genotyping, quantification
and imaging of gels, cloning, generating bacterial
strains and the central storage of bacteria,
vectors and cDNA constructs.
Equipment & Services Available
Any project requiring a large time commitment
by the core must be discussed with the Module
Director in advance.
In order to process projects more efficiently,
we ask that you provide the following written
information when requesting assistance from
the Molecular Biology Core:
For genotyping PCR
- -A short, written description of the
project and the name of the gene being
analyzed.
- -The manufacturer's data sheets for the
primers or the following information: primer
names, sequences, sizes, amount of primer,
and melting temperature. If the primers
have already been reconstituted, note their
concentration and diluent.
- -A protocol that includes cycling parameters
and magnesium concentration if available.
If a protocol is not available and conditions
need to be established by the core, please
allow time to determine the conditions.
i.e. Provide primers and "control" DNA
before tails/toes are snipped and ready
for analysis.
- -The expected product size for both wild
type and mutant/knockout PCR products.
- -The name, phone number, and e-mail address
of a contact person in your lab.
- -If you obtained mice from another laboratory,
a name, phone number, and e-mail address
for someone in that lab that has worked
with the mice.
For plasmid preparations and subcloning
projects
- -The complete plasmid name.
- -A map of the plasmid.
- -The plasmid's antibiotic resistance.
- -Any unusual conditions (such as special
bacteria or media) needed to grow the plasmid.
We routinely use LB. Advance notice is
needed to prepare other media.
Receiving plasmids from collaborators
When requesting clones from collaborators
for use in large scale plasmid preparations,
clones should be sent as follows (in order
of preference) whenever possible:
- -Purified DNA. One microliter of purified
DNA in a 0.5ml microfuge tube
or spotted on a small circle of Whatman
filter paper (0.5-1.0cm) is more than
adequate. DNA sent in this manner is
easily shipped, stored, and processed.
- -Streaked LB agar plates. Plates should
be streaked so that there are
individual, isolated colonies on the plate.
If sent from an institution that is not
in Cleveland, they should be shipped on
wet or artificial ice.
- -Stab cultures in vials.
- -Liquid cultures. These should be 24
hrs old or less. Liquid cultures
are accepted Monday through Thursday.
- -Dried/liquid bacteria and dried/liquid
DNA in pouches of plastic wrap
or aluminum foil are not ideal and should
be sent only as a last resort.
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