Date of Award

1988

Document Type

Thesis

Degree Name

Master of Science (MS)

Department

Biological Sciences

Program/Concentration

Biology

Committee Director

Andrew S. Gordon

Committee Member

Lloyd Wolfinbarger, Jr.

Committee Member

Paul Homsher

Call Number for Print

Special Collections LD4331.B47 S55

Abstract

A method was developed for extraction of plasmid DNA from bacterial cells embedded in agarose blocks. Cell - containing blocks were treated with various lysing reagents and inserted into the wells of an agarose gel. Upon electrophoresis the plasmid DNA migrated out of the embedding block and into the gel leaving intact chromosomal DNA in the well. This method was tested with various organisms and found to be effective for plasmid isolation. In comparison to "traditional" procedures, this new method is less tedious since chemical separation of plasmids is not required prior to electrophoresis. Also, a higher yield of plasmid DNA can be achieved due to the elimination of fractionation and precipitation steps involved in other methods. This method has the potential advantages of reduced shearing of DNA due to handling during isolation and, thus, may be useful in isolation of large plasmids and in studies which require that plasmids remain "supercoiled".

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DOI

10.25777/3v2h-vg67

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