Core Facilities: Molecular Biology

Susann Brady-Kalnay, Ph.D., Module Director
Department of Molecular Biology & Microbiology
Phone: (216) 368-0330
susann.brady-kalnay@case.edu

Moonkyung Hur Caprara, Ph.D., Module Manager
Department of Ophthalmology
Phone: (216) 368-4990
moonkyung.caprara@case.edu

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.

Forms for Download
Genotyping Request form.pdf
Cloning Request form.doc
Plasmid Prep form.pdf