65-90 Cloning a gene

65-90 Cloning a gene - Lab Recombinant DNA overview and...

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LAB Recombinant DNA: Ligation reaction Page # 65 Lab Recombinant DNA overview and ligation (Campbell, 7 th ed., pages 293-298, 384-388, 348-351. 8 th ed., pages 304-310, 396-400, 561-564 I Overview of constructing recombinant DNA (64-67) II. Ligation Reaction (67-68) Objectives: After completing labs two through six you should be able to: 1. Provide a general overview of how to construct recombinant DNA. 2. Explain the rationale for specific experimental manipulations. 3. Understand and be able to explain the general overview of constructing recombinant DNA. You should fully understand what a ligation reaction is and why you would perform one. Requirements: ( ) The pre-lab is due at the start of lab . The worksheet is due at the end of lab. Because the order of labs change each semester it is essential that you look at your detailed syllabus. The following material describes a typical cloning procedure. The initial overview is important to set the background for your actual experiments. Depending upon the semester you may or may not do the ligation reactions. You should still understand the theory behind the reactions even if you are not doing them. If you perform the ligation reaction you will continue up to Step 6 (page 68). You will then complete the rest of your lab exercises as listed in the syllabus. After which, you will return back to page 68 and finish Step 7, which is basically storing your samples until the next lab. Pages 69-88 will be addressed in future labs. All of the steps involved in constructing recombinant DNA are presented together on pages 64-88. Most students find in helpful to read all of these pages at this time so that you learn about cloning as a coherent unit. You will perform subsets of the cloning experiments in the next few labs. The summary diagram on page 64 will be very helpful. Realize that you may not be performing all of the steps shown in the summary diagram. In addition there is a glossary on pages 89 and 90. ( ) Constructing recombinant DNA. Biotechnology is a rapidly expanding area of science and industry. Concomitant with the growth of this field has been the development of new molecular tools to study DNA, gene expression, protein production, protein shape, etc. In spite of these advances the simple technique of constructing recombinant DNA is still essential. You will learn most of the techniques required to construct recombinant DNA. Recombinant DNA refers to a DNA molecule that contains DNA from two different DNA sources (usually this is from two different species or even DNA from non-living organisms such as viruses and plasmids). The theory of constructing recombinant DNA is not that difficult and it will be discussed in greater detail in lecture and in later labs. In the next few labs you will be given the minimal theory necessary to slightly understand the experimental steps. We are doing this because this will allow us to follow approximately the normal chronological steps of constructing recombinant DNA.
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65-90 Cloning a gene - Lab Recombinant DNA overview and...

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