Director: Andrew Stolz, M.D.
Co-Director: Juergen Reichardt, Ph.D.
Research Assoc: Shou Liu
Research Tech: Maria Villalvazo
Contact phone number: Core: (323) 442-2164, FAX: (323) 442-3420

Core Services

A) Oligonucleotide Synthesis Facility:

Core will synthesis oligonucleotides at a cost of $ 1.25 per base for unpurified oligonucleotide with an anticipated turnover of 48 to 72 hours. In the future, Core may provide phosphothioamidated or biotinylated primers based on demand..

B) Isolation and Preparation of Xenopus oocytes for Microinjection Studies:

Molecular Biology Core will establish and maintain a colony of frogs for Xenopus oocyte isolation required for transport and cloning studies. We anticipate a weekly (Monday) preparation of oocytes with a charge of $ 25 per ovary. We will harvest and provide investigators will a pool of enriched oocytes suitable for expression studies. Individual investigators will be responsible for subsequent experiments such as microinjection or functional assay with oocytes. A microinjection station is available for use and training.

C) Plasmid Preparation:

Core will isolate on a weekly schedule plasmids containing DNA of interest at a cost of $15 per 150 ml of bacterial culture. Plasmid will be harvested using column chromatography (Qiagen) with potential for cesium chloride purification.

D) Custom Services:

1) Recombinant Protein Expression in Bacteria:

Soluble proteins or peptide fragments will be expressed in bacteria as either a fusion protein or by itself. Fusion protein will be isolated by Core and provided to investigators for subsequent studies. Charge for service will be established to cover supplies based on protocol and amount of protein desired.

2) Basic Subcloning:

Core will provide basic subcloning for investigators who are currently not using standard molecular biological techniques. This service can be used to generate riboprobes for hybridization studies, constructs for bacterial expression studies and subcloning of cDNA for DNA sequencing. Cost will be established to cover supplies.

The following equipment and teaching services will be provided by the Molecular Biology Core Service.

A) Equipment:

All equipment will be available by using a reservation time sheet which will be used to document usage of equipment.

1) Molecular Biology Data Analysis:

A Macintosh PowerPC work station is available for DNA sequence analysis with the following capacity: restriction mapping, alignment of contiguous sequences, optimum sequence for PCR primers, entering DNA sequences from DNA sequencing gels, identification of open reading frames and proteolytic cleavage restriction mapping. Comparison of DNA or protein sequences with the most current data bases will be performed by Blast program available by the Internet. Guidance will be provided in using these different programs and in the analysis of results.

2) Xenopus Oocyte Microinjector:

A fully functional variable delivery Drummond Microinjection is available for Xenopus oocyte studies and other uses. Instruction will also be available for pulling of microinjector needles and injection of oocytes.

3) Core will also be used as a learning center for Center members who desire to incorporate molecular biological techniques into their current research program. Technician and Research Associate can be trained in nucleic acid isolation, standard blotting and hybridization techniques, RN'ase protection and DNA sequencing. For Center members, individual consultation with Core directors will also be available.