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Related Experiment Video

Updated: Jul 5, 2026

Generating Transgenic Plants with Single-copy Insertions Using BIBAC-GW Binary Vector

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Construction of YAC libraries with large inserts.

Simon Foote¹, Christopher Denny

¹Whitehead Institute, Cambridge, Massachusetts, USA.

Current Protocols in Human Genetics

|April 23, 2008

Summary

This study details the yeast artificial chromosome (YAC) cloning system for large genomic DNA fragments. It outlines protocols for YAC vector preparation, DNA transformation into yeast spheroplasts, and methods for partial genomic DNA digestion.

Area of Science:

Molecular Biology
Genomics
Biotechnology

Background:

The yeast artificial chromosome (YAC) system enables cloning of large genomic DNA fragments.
Libraries with megabase-pair inserts have been generated using YAC technology.

Purpose of the Study:

To describe the protocol for preparing YAC vectors.
To outline the transformation of ligated DNA into yeast spheroplasts.
To detail methods for partial genomic DNA digestion.

Main Methods:

Preparation of YAC vectors.
Transformation of ligated DNA into yeast spheroplasts.
Two methods for partial genomic DNA digestion: EcoRI restriction endonuclease-EcoRI methylase competition and Mg2+-limited digestion.

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Main Results:

Successful preparation of YAC vectors.
Efficient transformation of ligated DNA into yeast spheroplasts.
Established protocols for partial genomic DNA digestion.

Conclusions:

The yeast artificial chromosome system is effective for cloning large genomic DNA.
Detailed protocols facilitate YAC library construction and genomic DNA manipulation.