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ANALYSIS OF RECOMBINANT DNA USING THE ALKALINE LYSIS METHOD.
Anyanna S. Flemings,Kisha Vanterpool, lisa Williams, Marc Alston; mentor, Dr. Matthew George
Department of Biochemistry.
Within the past two decades it has become a common pratice to combine DNA sequence from
different sources. This process has been known as Recombinant Techonology. Creating a recombinant
DNA molecule from two different sources allows for the DNA fragment of intrest to be cloned when
inserted into host cells, such as E. coli. Iisertion of foregin DNA into host cells is refered to
as transformation. When the recombinant molecule is placed in cells such as bacteria, the molecule is able to replicate independently of the host c chromosome when it is grown in the appropriate
medium. Therefore, when isolated, large amounts of the recombinant molecule can be produce and
used for further studies. We studied a recombinant DNA molecule which contained a Polymerase
Chain Reaction (PCR) generated from hair samples, amplified by PCR, and cloned into E. coli
The objective fo this experiment is to characterize or confirm the presence of 400 base pair
mitochondial DNA (mt DNA) fragment which had been cloned into E. coli cells. In order to
characterize the reconmbinant molecule, we first had to isolate the recombinant DNA. This was
performed by the Alkaline Lysis Method. The recombinant DNA was islolated, digested with a
restriction enzyme, and electrophorsed on an agarose gel.
Our result showed that we had recovered the 400 base pair mt DNA fragment from the recombinant
molecule.
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Anyanna S. Flemings, a May 1996 Howard University graduate, plans to attend graduate
school at Howard University, where she will earn a doctorate in biology. As an undergraduate,
Flemmings undertook a rigorous academic curriculum while working two jobs and participating in a
number of extracurricular activities. She was a member of the NAACP, the Health Professions Club,
and the Biological Science Society. She was also a resident assistant and a summer assistant. i
After earning her doctorate, Flemmings plans to teach. "I would like to be that constant vehicle
of encouragement for young, disadvantaged, low-income minotities," whom she say she can relate to
personally. A biology major and chemistry minor, Flemmings is a native of Roseville, Michigan. In recommending her for the McNair Program, one of her professor said he expected Flemmings to one
day be a true leader in her field.
McNair Alumni
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