International Mammalian Genome Society


The 13th International Mouse Genome Conference
October 31-November 3, 1999

Table of Contents * Structure * Bioinformatics * Sequence * Mapping * New Tools * Gene Discovery * Developmental * Mutagenesis * Functional Genomics

D12 Archiving and Recovery of Bacterial Plasmids Using FTA Paper

Michael Connolly, Pamela Hansen, Arubula Reddy, and Mindy Goldsborough. Research and Development, Life Technologies Inc., 9800 Medical Center Dr., Rockville, MD USA 20850

The generation of bacterial plasmids/clones is a common protocol in many laboratories. Numerous genome projects are currently underway in which large quantities of clones are being generated. In order to ensure project continuity, creating archives of the clones generated is an integral part of the process. However, archiving these clones using glycerol storage at -80oC requires vast amounts of freezer space. FTA is a new storage/archiving system that allows for the long-term storage of DNA at room temperature in a compact format.

FTA paper is a cellulose-based matrix impregnated with chemicals that lyse cells but also protects and stabilizes DNA for long-term room temperature storage. This medium has been used for several years in the Forensic field for the collection and storage of blood samples. Blood preserved on FTA has been shown to remain amenable to PCR for at least 10 years.

It has also been demonstrated that bacterial cells from an overnight culture are easily lysed on FTA paper. Furthermore, it has been shown that samples prepared in this manner are stable for long periods of time. We will present data on the archiving and recovery of bacterial plasmid clones on FTA paper. Bacterial samples can be applied to FTA in several forms: resuspended colonies, overnight cultures, glycerol stocks or purified DNA.

Plasmid clones are recovered from FTA by transformation; a small "punch" of sample is added to competent cells. We have successfully archived and recovered plasmids from 2 Kb to 227 Kb (BACs). To facilitate the use of FTA in large genome projects, FTA is available is a 96 well robot compatible format.

Other uses of FTA that will be discussed include the direct sequencing of M13 and bacterial cultures stored on the paper and the use of FTA as a simple clone distribution method.

 


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