Title

Genomic DNA microextraction: a method to screen numerous samples

UMMS Affiliation

Department of Cell Biology

Date

3-1-1992

Document Type

Article

Medical Subject Headings

Animals; Blotting, Southern; Clone Cells; DNA; Genome; Humans; Mice; Restriction Mapping; Stem Cells

Disciplines

Cell Biology

Abstract

Many experimental designs require the analysis of genomic DNA from a large number of samples. Although the polymerase chain reaction (PCR) can be used, the Southern blot is preferred for many assays because of its inherent reliability. The rapid acceptance of PCR, despite a significant rate of false positive/negative results, is partly due to the disadvantages of the sample preparation process for Southern blot analysis. We have devised a rapid protocol to extract high-molecular-weight genomic DNA from a large number of samples. It involves the use of a single 96-well tissue culture dish to carry out all the steps of the sample preparation. This, coupled with the use of a multichannel pipette, facilitates the simultaneous analysis of multiple samples. The procedure may be automated since no centrifugation, mixing, or transferring of the samples is necessary. The method has been used to screen embryonic stem cell clones for the presence of targeted mutations at the Hox-2.6 locus and to obtain data from human blood.

Rights and Permissions

Citation: Anal Biochem. 1992 Mar;201(2):331-5.

Related Resources

Link to Article in PubMed