Title

Direct Usage of Taq Polymerase in Live E. coli for PCR

Poster Number

19

Lead Author Major

Biochemistry/Biological Sciences/Chemistry

Format

Poster Presentation

Faculty Mentor Name

Craig Vierra

Faculty Mentor Department

Biological Sciences

Abstract/Artist Statement

Taq polymerase is an essential enzyme and commonly used in PCR reactions. DNA polymerase is isolated from the bacteria Thermus aquaticus, which lives in hot springs, giving the enzyme heat stability with an optimum temperature around 75-80 °C. This allows Taq to elongate the target DNA fragments at high temperatures without denaturation. To date, most labs require purification of Taq polymerase from E. coli for PCR reactions. This process can be time consuming and expensive. The purpose of our studies was to find a faster, more convenient method to perform PCR without the need to purify Taq polymerase from bacteria. In our project, we attempted to run PCR reactions using intact bacterial cells that were expressing Taq as our source of DNA polymerase. Here we demonstrate that PCR can be successfully performed without the need to purify Taq polymerase from bacteria. We show that intact bacterial cells expressing Taq polymerase can be lysed directly during the initial steps of PCR, providing a functional DNA polymerase for amplifying DNA templates. Lastly, we demonstrated that the bacterial cells can be stored and subject to freeze thawing without the loss of DNA polymerase activity. This enables lab researchers to have more efficient, convenient, and economic way to use Taq polymerase in PCR reactions. This technique has been successfully proven, and can be applied in a large variety of lab research.

Location

DeRosa University Center, Ballroom

Start Date

21-4-2011 6:00 PM

End Date

21-4-2011 8:00 PM

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Apr 21st, 6:00 PM Apr 21st, 8:00 PM

Direct Usage of Taq Polymerase in Live E. coli for PCR

DeRosa University Center, Ballroom

Taq polymerase is an essential enzyme and commonly used in PCR reactions. DNA polymerase is isolated from the bacteria Thermus aquaticus, which lives in hot springs, giving the enzyme heat stability with an optimum temperature around 75-80 °C. This allows Taq to elongate the target DNA fragments at high temperatures without denaturation. To date, most labs require purification of Taq polymerase from E. coli for PCR reactions. This process can be time consuming and expensive. The purpose of our studies was to find a faster, more convenient method to perform PCR without the need to purify Taq polymerase from bacteria. In our project, we attempted to run PCR reactions using intact bacterial cells that were expressing Taq as our source of DNA polymerase. Here we demonstrate that PCR can be successfully performed without the need to purify Taq polymerase from bacteria. We show that intact bacterial cells expressing Taq polymerase can be lysed directly during the initial steps of PCR, providing a functional DNA polymerase for amplifying DNA templates. Lastly, we demonstrated that the bacterial cells can be stored and subject to freeze thawing without the loss of DNA polymerase activity. This enables lab researchers to have more efficient, convenient, and economic way to use Taq polymerase in PCR reactions. This technique has been successfully proven, and can be applied in a large variety of lab research.