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High throughput direct end sequencing of BAC clones
J M Kelley1, C E Field, M B Craven
1The Institute for Genomic Research, Rockville, MD 20850, USA and Division of Biology,California Institute of Technology, Pasadena, CA 91125, USA.
Nucleic Acids Research
|February 26, 1999
Summary
We developed a low-cost, high-throughput method for bacterial artificial chromosome (BAC) end sequencing. This technique enables efficient selection of minimally overlapping clones for large-scale genomic sequencing projects.
Area of Science:
- Genomics
- Molecular Biology
- Biotechnology
Background:
- Bacterial artificial chromosome (BAC) libraries are crucial for high-throughput genomic sequencing due to their stability.
- BAC end sequences are proposed for selecting minimally overlapping clones for sequencing large genomic regions.
- Developing high-throughput BAC end sequencing methods is essential for this strategy's effectiveness.
Purpose of the Study:
- To describe a cost-effective and efficient 96-well procedure for BAC end sequencing.
- To enable the generation of BAC end sequences from deep coverage BAC libraries.
Main Methods:
- A low-cost, 96-well procedure for BAC end sequencing was developed.
- The method was applied to human and Arabidopsis BAC libraries.
Main Results:
- Generated BAC end sequences with an average read length greater than 450 bases.
- Achieved a single-pass sequencing accuracy greater than 98% for human and Arabidopsis libraries.
- Demonstrated the efficiency and cost-effectiveness of the developed BAC end sequencing method.
Conclusions:
- The developed BAC end sequencing method is efficient and cost-effective.
- This method facilitates the selection of minimally overlapping clones for large-scale genomic sequencing.
- The application of BAC end sequences is vital for advancing genomic sequencing projects.