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Amplification of cosmid and plasmid libraries.
1Harvard Medical School, Boston, Massachusetts, USA.
Current Protocols in Molecular Biology
|February 12, 2008
Summary
This study details a protocol for amplifying bacterial reagent libraries, enabling multiple uses from a single preparation. Storing amplified cosmid or plasmid libraries in glycerol allows for over 1000 platings from a small volume.
Area of Science:
- Molecular Biology
- Microbiology
Background:
- Reagent libraries are essential for various molecular biology applications.
- Efficient amplification protocols are needed to maximize the utility of these libraries.
Purpose of the Study:
- To describe a protocol for amplifying bacterial reagent libraries.
- To enable repeated use of cosmid or plasmid libraries.
Main Methods:
- Bacteria with recombinant clones are cultured on agar plates.
- Bacterial cells are harvested and stored in glycerol.
- Amplified libraries are prepared in small volumes (10-100 microliters).
Main Results:
- An amplified library provides one library's equivalence in a small volume.
- Storing amplified libraries in glycerol facilitates long-term preservation.
- A 100 microliter amplified library can yield over 1000 platings.
Conclusions:
- This protocol offers a method for creating stable, reusable reagent libraries.
- Amplification significantly increases the number of experimental applications possible from a single library.
- The glycerol storage method ensures library viability for extensive downstream use.
