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Size fractionation using agarose gels.

T Quertermous¹

¹Massachusetts General Hospital, Boston, Massachusetts, USA.

Current Protocols in Molecular Biology

|February 12, 2008

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This study details methods for size-fractionating genomic DNA using agarose gel electrophoresis. These techniques enable the purification of large DNA amounts for constructing genomic libraries and generating numerous recombinant clones.

Area of Science:

Molecular Biology
Genomics
Biotechnology

Background:

Genomic library construction requires purified, size-fractionated DNA.
Traditional methods can be limiting for large-scale DNA purification.

Purpose of the Study:

To present optimized protocols for size-fractionating digested genomic DNA.
To facilitate the creation of high-quality genomic libraries.

Main Methods:

Basic Protocol: Slab agarose gel electrophoresis, Southern blot analysis for identification, and DNA elution.
Alternate Protocol: Preparative circular agarose gel electrophoresis using a Bull's-eye apparatus for DNA fractionation.

Main Results:

Successful purification of large quantities of size-fractionated genomic DNA.

Related Experiment Videos

Identification of target DNA fractions using Southern blotting.

Demonstrated suitability for genomic library construction.

Conclusions:

The described protocols efficiently yield size-fractionated DNA suitable for genomic library construction.
These methods support the generation of a large number of recombinant clones.