Biotechnology: Analytical Techniques

by Kevin Ahern, PhD

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    It is said that knowledge is power and with biotechnology we can see in this presentation how it is true where the knowledge about molecular biological processes has been brought to develop powerful analytical techniques. I am going to talk about three of those techniques in this presentation. DNA amplification, microarray analysis and 2D gel analysis. Now, knowledge about the process of DNA amplification has been applied in a technique called the polymerized chain reaction that has revolutionized the way that we see and analyze DNA. The polymerized chain reaction allows a person to amplify specific DNA segments out of a larger DNA millions of times. It's very popular for a forensic analysis and if you ever watched a crime show on TV you have probably seen PCR being applied to action. The process of PCR or polymerized chain reaction is derived simply by copying some ideas from cellular DNA replication and then using this in a noble way to accomplish the replication of DNA that one desires only to copy in a specific way. The advantage of PCR is that it is a very simple technique to perform A high school student can be trained to do PCR in about an hour. It requires sequence knowledge to start and that's one of the primary requirements of PCR, as we shall see. Now the PCR technique uses the knowledge of the sequence to design and chemically synthesize DNA primers flanking the region to be amplified. Now I need to explain that. In a previous lecture I talked about how DNA polymerase uses a primer to start DNA replication and in the cell that primer is RNA. Now the problem with the RNA primer is, it’s got to be removed and then replaced by something else. That's kind of a complicated...

    About the Lecture

    The lecture Biotechnology: Analytical Techniques by Kevin Ahern, PhD is from the course Analytical Techniques in Biotechnology. It contains the following chapters:

    • DNA Amplification
    • Microarray Analysis
    • Proteomics - 2D Gel Analysis

    Included Quiz Questions

    1. ...has three steps to each cycle.
    2. ...requires an RNA primer for each round of replication.
    3. ...works best with a heat-sensitive DNA polymerase.
    4. ...typically amplifies DNA about 1000 fold.
    1. ...denaturation is done first.
    2. ...renaturation must be done very quickly.
    3. ...elongation must be done at temperatures near boiling.
    4. ...actual doubling of the amount of DNA occurs with each cycle.
    1. ...can be performed with microarray analysis.
    2. ...tells how much protein is in a cell.
    3. ...focuses mostly on rRNA.
    4. ...focuses mostly on tRNA.
    1. RNA fragments are attached to a grid.
    2. DNA fragments are attached to a grid.
    3. Annealing of RNAs to the DNA is the key step in the process.
    4. mRNAs are the RNAs being analyzed.
    1. ...each spot on the final gel corresponds to a single protein.
    2. ...the first step involves separating proteins by molecular weight.
    3. ...SDS is necessary to separate proteins by pI.
    4. ...the smallest proteins are found at the top of the gel.

    Author of lecture Biotechnology: Analytical Techniques

     Kevin Ahern, PhD

    Kevin Ahern, PhD

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