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Module 3-BSC2010L
BDC2010L- DNA Extraction, The PRC reaction, DNA Electrop, Bacteria Transf
| Question | Answer |
|---|---|
| What does PCR stand for? | Polymerase chain reaction |
| DNA in the sub-mini unit moves? | From negative (left/black) to positive (right/red) |
| PCR is dependent on a heat-stable | Polymerase. The heat-stable DNA polymerase is used to extend the primers after they have annealed to the template DNA. |
| At which of the following temperatures would you predict maximum activity from a heat-stable polymerase such as Taq polymerase? | 72oC 72oC is the optimal temperature for many heat-stable polymerases. |
| Place the steps of the Polymerase Chain Reaction in the correct sequence. | Denaturing, Annealing, Extending (replicating) The template DNA is separated into single strands (denaturing) the primers bind to their complementary sequence on the template (annealing), a heat-stable polymerase extends the primers (replicating). |
| The denaturing and annealing steps of PCR involve the breaking or formation of _____ bonds. | hydrogen Complementary base pairing, between template strands or between primers and template, is based upon hydrogen bonds between nitrogenous bases of a base pair. |
| he Polymerase Chain Reaction is used to ________a specific sequence of DNA. | amplify PCR is used to make copies of, or amplify, a specific sequence of DNA. |
| Which of the following is used to detect the products of PCR? | Gel electrophoresis PCR products can be detected and analyzed using gel electrophoresis. |
| Please select the statement that best describes the purpose of gel electrophoresis. | Gel electrophoresis separates DNA fragments based on size. Gel electrophoresis is able to separate DNA fragments based on their size. The smaller fragments travel the most quickly through the gel and will be the farthest from the wells. The larger fragme |
| In the gel electrophoresis lab, you determined the ________ of three different DNA samples. | genotype |
| Gel electrophoresis is an important tool used in biotechnology for forensics and paternity test. | True |
| How does the DNA rate of travel differ for small DNA fragments and large DNA fragments? | Small fragments travel farther than large fragments. |
| Put these DNA fragments in order of their position after gel electrophoresis, starting with the one closest to the negative end of the gel and ending with the one closest to the positive end of the gel. | 100 bp, 73 bp, 42 bp, 25bp The largest fragment would move the slowest and would remain closest to the negative end where it started in the well. The smallest fragment would move the fastest and be located closest to the positive pole. |
| In gel electrophoresis, how are the DNA samples added to the wells? | A micropipet is used. |
| How does the size of a DNA fragment relate to its speed of passage through the agarose gel? | Smaller fragments move through the gel faster. Agarose is added to solidify the gel. It then acts as a filter, allowing only smaller particles to pass through quickly. Larger molecules are slowed by this and do not move as fast or far through the gel. |
| Why are DNA fragments attracted to the positive pole of the agarose gel? | DNA is negatively charged. DNA is a negatively charged molecule, and therefore, it is attracted to the positive pole of the gel. |
| During the process of transformation in the lab setting, cells are plated on selective media to: | make sure that only transformants grow |
| Cells that are capable of bringing DNA from their environment in through their cell wall are called: | competent |
| Transformation is facilitated by: | proteins on the cell wall that bind DNA from the environment |
| The acquisition of genetic material through the uptake of “naked” DNA molecules from the environment by bacteria is called: | Transformation |
| encodes resistance to ampicillin | bla |
| DNA polymerase begins replicating the plasmid at this site | ori |
| regulatory protein that affects the transcription of the gene for green fluorescent protein | araC |
| gene that encodes green fluorescent protein | GFP |
| What is the role of arabinose in the transformation procedure? | It is an inducing substrate that allows the transcription of the gene of interest. |
| After incubation, which plate is likely to have zero growth? | -pGLO in LB/amp |
| What parts of the transformation procedure are done to increase the competence of the bacterial cells? | treatment with calcium, brief heat shock |
| Which plate(s) is/are selective for the transformed bacterial cells? | LB/amp agar, LB/amp/ara agar |
| What is the role of arabinose in the transformation procedure? | green fluorescence under UV light, ampicillin resistance |
| Why is it important to keep the filtrate and reagents cold? | Cold temperature stabilizes DNA / inhibits DNA breakage |
| What is the purpose of mixing cells with NaCl and detergent to form the strawberry filtrate? | The detergent emulsifies cell and nuclear membranes of cells by disrupting the phospholipid bilayers. The sodium ions neutralize the negative charge of DNA to facilitate precipitation. This helps the DNA come out of the solution. |
| What is the purpose of the meat tenderizer? | To digest associated proteins that bind the DNA. Proteins help organize DNA into a packed structure inside a cell, so these need to first be removed. |
| What is the purpose of the ice cold 95% ethanol? | To precipitate the DNA from the filtrate. DNA is not soluble in ethanol but it is in the water-based filtrate. As a result, the DNA precipitates when it enters the ethanol allowing it to be spooled. |
| What is the correct wat to spool the DNA? | Slowly, using a constant motion and pressure to collect the precipitate. This gives the best chance at collecting intact DNA strands. |
| Steps to isolate DNA | 1. Place test tube on ice 2. Add strawberry filtrate to test tube. 3. In a graduated cylinder, dissolve meat tenderizer with distilled water. 4. Add meat tenderizer to strawberry filtrate , let it react for 10 min. 5. add 95% ethanol to precipitate |
| Select all of the following that are components of DNA | Nitrogenous base, phosphate group and deoxyribose |
| What is the product of DNA replication? | Two daughter DNA molecules |
| The process of forming RNA from a template strand of DNA is called _____. | Transctiption |
| After DNA replication, each new daughter DNA molecule is composed of ________________. | one parental strand and one newly synthesized strand |
| In transcription, the enzyme RNA polymerase builds a strand of RNA along the __________. | template strand of DNA |
| The 2 strands of nucleotides in DNA molecules are held together by _____________. | hydrogen bonds between base pairs |
| What is the purpose of installing the comb? | to create a well /pockets in the gel where samples can later be placed |
| Why is ethidium bromide added at this stop? | Ethidium bromide is needed to see the DNA bands in the gel under UV illumination. |
| Why is it important to pour slowly and avoid air bubbles? | Bubbles interfere with the movement of the sample through the gel, distorting the results. |
| What would happen if the gel was run for too long? | The sample bands would move too far and leave the bottom of the gel. |
| Ori site | location for DNA polymerase to start replication of the plasmid |
| Bla gene | confers resistance to ampicillin, allowing transformed cell and its progeny to survive on media containing ampicillin |
| GFP gene | taken from jellyfish, it encodes for a green fluorescent protein, the gene of interest in this simulation |
| araC gene | regulates transcription of the GFP gene, which is active in the presence of arabinose |
| What is the purpose of the LB agar plate? | it is a selective medium for bacterial growth |
| What is the purpose of the LB/amp agar plate? | it is a selective medium containing an antibiotic inhibit the growth of non-resistant cells |
| What is the purpose of the LB/amp/ara agar plate? | it is a selective medium with an antibiotic which prevents growth of non-resistant cells and a sugar that allows for the expression of a plasmid gene. |
| What steps were taken to increase the efficiency of the transformation process? | Treatment with calcium chloride and brief heat shock |
| What do you need for PCR? | thermostable DNA polymerase, Taq polymerase, and requires DNA primers designed specifically for the DNA region of interest, nucleotides and an enzyme |
| What do you need for PCR? | Taq polymerase, primers, template DNA, and nucleotides (DNA building blocks). The ingredients are assembled in a tube, along with cofactors needed by the enzyme, and are put through repeated cycles of heating and cooling that allow DNA to be synthesized. |
| Natural selection can alter the frequency distribution of heritable traits in three ways, depending on which phenotypes in a population are favored: _______________________, _____________, and _______________. | through directional selection, disruptive selection, and stabilizing selection. |
| What is Directional selection? | When conditions favor individuals exhibiting one extreme of a phenotypic range, thereby shifting a population’s frequency curve for the phenotypic character in one direction or the other. |
| What is Disruptive selection? | When conditions favor individuals at both extremes of a phenotypic range over individuals with intermediate phenotypes |
| What is Stabilizing selection? | It acts against both extreme phenotypes and favors intermediate variants. This mode of selection reduces variation and tends to maintain the status quo for a particular phenotypic character. |
| What is Intersexual selection? | Also called mate choice, individuals of one sex (usually the females) are choosy in selecting their mates from the other sex. |
| What is frequency-dependent selection? | The fitness of a phenotype depends on how common it is in the population. |
| List 4 reasons why Natural Selection Cannot Fashion Perfect Organisms. | Selection can act only on existing variations. Evolution is limited by historical constraints. Adaptations are often compromises. Chance, natural selection, and the environment interact. |
Created by:
Jen2pen
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