DNA Isolation and Insertion

DNA Isolation and Insertion - Biochemistry Lab December 2...

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Biochemistry Lab December 2, 2005 DNA Isolation and Insertion into a Vector with Restriction Digest are Examined on an Agarose Gel Objectives: Vector insertion is a useful tool in research. Techniques have been used which can insert a target DNA into bacterial DNA. The circular DNA is nicked and the target DNA is then annealed to the bacterial DNA and then the circle is re-closed. This allows scientists to hi-jack the bacteria’s own DNA replication to create huge numbers of copies of their target DNA. The DNA can then be extracted and digested and separated on a gel. The target DNA can then be determined by size on the gel and cut out of the agarose and purified. This purified DNA is your inserted vector and can be sequenced for analysis. Procedure: First we gathered some cells and centrifuged them to create a pellet. The we added 100 uL LYR solution and resuspended the cells by pipetting up and down. The mixture was then incubated for 5-10 minutes at room temperature. Next we added 200
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DNA Isolation and Insertion - Biochemistry Lab December 2...

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