Molecular Engineering
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Molecular Engineering
- 1. Central Dogma of DNA -Marif lor P. Ramos
- 2. + (space) -specta + -k = ?
- 4. What Is MOLECULAR GENETICS? Molecular genetics is the study of the agents that pass information from generation to generation. These molecules, our genes, are long polymers of deoxyribonucleic acid, or DNA. Just four chemical building blocks guanine (G), adenine (A), thymine (T), and cytosine (C)—are placed in a unique order to code for all of the genes in all living organisms.
- 5. GUANINE CYTOSINE
- 6. ADENINE THYMINE
- 7. Genes determine hereditary traits, such as the color of our hair or our eyes. They do this by providing instructions for how every activity in every cell of our body should be carried out. For example, a gene may tell a liver cell to remove excess cholesterol from our bloodstream. It will instruct the cell to make a particular protein. It is this protein that then carries out the actual work. Many diseases are caused by mutations, or changes in the DNA sequence of a gene. When the information coded for by a gene changes, the resulting protein may not function properly or may not even be made at all. In either case, the cells containing that genetic change may no longer perform as expected.
- 8. Isolating DNA from just a single cell provides a complete set of all a person's genes, that is, two copies of each gene. However, many laboratory techniques require that a researcher have access to hundreds of thousands of copies of a particular gene. One way to obtain this many copies is to isolate DNA from millions of cells grown artificially in the laboratory. Another method, called cloning, uses DNA manipulation procedures to produce multiple copies of a single gene or segment of DNA. The polymerase chain reaction (PCR) is a third method whereby a specific sequence within a double-stranded DNA is copied, or amplified. PCR amplification has become an indispensable tool in a great variety of applications.
- 9. Cell culture involves growing cells under artificial conditions, either attached to some type of artificial surface or suspended in a special solution. In both cases, the cells are bathed in fluids containing nutrients that are either synthetically produced or extracted from related organisms. Certain cell types are more amenable to being grown in culture than others. Conditions that serve to sustain one cell type may not apply to other cell types, or even the same cell type from another species. Cell culture is a useful technique because it provides a renewable source of cells for isolating DNA.
- 10. DNA isolation refers to the process of extracting DNA from a cell in a relatively pure form. It involves separating DNA from other cellular components, such as proteins, RNA, and lipids. Whatever the source, the DNA is isolated by placing the cells in a tube containing a special solution, called a "cocktail", and mechanically or chemically breaking them open. This causes the cell to release its contents into the cocktail containing enzymes, chemicals, and salts. Enzymes are used to chew up the proteins; chemicals to destroy any RNA present; and salts to help pull the DNA out of solution. At this point, the DNA will exist in long strands that form a mucous-like glob within the solution. The DNA is then harvested by spinning the tube in a machine called a centrifuge. During spinning, the DNA collects in the bottom of the tube. The solution is then poured off, and the DNA is dissolved in a second solution that will make it easy to work with in subsequent procedures. The result is a concentrated DNA sample containing many thousands of copies of each gene.
- 11. In the first step of mRNA isolation, a cell is ruptured, and the cellular contents are exposed to synthetic beads coated with strings of thymine nucleotides. Because adenine and thymine readily bind to each other, poly(A) mRNA is selectively retained on the beads while the other cellular components are washed away. Once isolated, purified mRNA is converted to single-stranded DNA using the enzyme reverse transcriptase and is then made into a stable double-stranded DNA using the enzyme DNA polymerase. DNA produced in this way is called complementary DNA (cDNA) because its sequence, at least the first strand, is complementary to that of the mRNA from which it was made.
- 12. mRNA Isolation This drawing demonstrates how poly(A) RNA can be isolated from other RNAs by separation on a special solid support material. In this example, the material is made up of glass beads to which thymine molecules are attached. Because adenine and thymine molecules readily bind to each other, mRNAs with poly(A) tails will be selectively retained on the beads. As seen on the left-hand side of the diagram, a solution containing various RNA populations, including mRNAs with poly(A) tails (red) as well as other RNAs and cellular material (purple), is applied to the separation column. Only the poly(A) RNA is retained, because it is immobilized on the solid support material. The other RNAs and cellular material pass through the column. On the right, the bound poly(A) mRNA is retrieved by treating the column with a special buffer solution that breaks the thymine nucleotide–AAA bond. The mRNA can be collected in a tube for further experimentation.
- 13. The porous and thin nature of a gel is ideal for separating DNA fragments using electrophoresis, but as we mentioned earlier, these gels are delicate and rarely usable for other techniques. For this reason, DNA that has been separated by electrophoresis is transferred from a gel to an easy-to-handle inert membrane, a process called blotting. Term "blotting" describes the overlaying of the membrane on the gel and the application of a pad to ensure even contact, without disturbing the positions of the DNA fragments. This procedure reproduces the exact pattern of DNA captured in the gel on the membrane. The membrane can then be probed with a DNA marker to verify the presence of a target sequence.
- 14. The process of determining the order of the nucleotide bases along a DNA strand is called sequencing. In 1977, 24 years after the discovery of the structure of DNA, two separate methods for sequencing DNA were developed: the chain termination method and the chemical degradation method. Both methods were equally popular to begin with, but, for many reasons, the chain termination method is the method more commonly used today. This method is based on the principle that single-stranded DNA molecules that differ in length by just a single nucleotide can be separated from one another using polyacrylamide gel electrophoresis, described earlier.
- 16. Answer the following What is What questions. Molecular happens Genetics? during RNA When you Isolation? already saw the picture, give their What is Give the four essence to Cell building Human Culture? blocks. Life. What Is CLONNING?
- 17. Stick It On! Translate the Question using the STICKS Graph. Then, well of course, ANSWER IT. J K L S W A B C T U X Y D E F M N O V Z G H I P Q R QUESTION: ?