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The construction and analysis of M13 libraries prepared from YAC DNA
M Vaudin1, A Roopra, L Hillier
1Department of Genetics, Washington University School of Medicine, St. Louis, MO 63108.
Nucleic Acids Research
|February 25, 1995
Summary
This study details methods for creating high-quality shotgun libraries from yeast artificial chromosome (YAC) DNA. Survey sequencing provides a rapid way to characterize unknown YAC and cosmid clones.
Area of Science:
- Genomics
- Molecular Biology
- Bioinformatics
Background:
- Yeast artificial chromosomes (YACs) are crucial for isolating and mapping large genomic DNA segments.
- Extensive use of YACs in genome analysis necessitates efficient DNA handling and characterization methods.
Purpose of the Study:
- To develop and refine procedures for generating high-quality shotgun libraries from limited YAC DNA.
- To assess the utility of survey sequencing for rapid characterization of YAC and cosmid clones.
Main Methods:
- Modification of existing procedures for producing shotgun libraries from YAC DNA.
- Sequencing of clones from multiple libraries.
- Analysis of sequence distribution, integrity, and yeast DNA contamination.
- Database comparison of sequences (survey sequencing).
Main Results:
- Successful generation of high-quality shotgun libraries from small amounts of YAC DNA.
- Analysis confirmed sequence integrity and manageable levels of yeast DNA contamination.
- Survey sequencing at 1-fold coverage proved effective for characterizing unknown clones.
Conclusions:
- The described procedures enable efficient creation of YAC DNA libraries.
- Survey sequencing is a fast and reliable method for preliminary characterization of cosmid and YAC clones.