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20 Cards in this Set
- Front
- Back
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What is Form II DNA?
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nicked DNA that relaxes into a circle
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How is DNA nicked?
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break the phosphate bonds in supercoiled DNA using DNase or mechanically
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What are the names of the interlocking rings that plasmids form during replication?
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Catenane
Dimer (2) Trimer (3) |
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How is the alkaninity of lysate solution neutralized?
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Potassium acetate neutralization buffer
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What does PRC do?
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It is a technique that enables laboratory amplification of large amounts of a particular segment of DNA, even if it is in a complex mixture of other DNA.
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What is "real time" PCR?
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Real time PCR allows researchers to quantify reactions by monitoring the amount of DNA produced during amplification.
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How do we determine the amount of biological material during PCR?
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If the amount pf product produced is proportional to the initial amount of DNA amplified, then we can extrapolate the biological quantities in cells through amplification.
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How do we quantify real-time PRC?
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It uses interworking florescent dye markers and quenchers that work within the process. The amount of light emitted by the dye is a quantitative indicator of how much new DNA had been produced.
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What is reverse transcriptase?
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It is an enzyme that detects the presence of minute amounts messernger RNA present in a cell and converts into mRNA into DNA that can then be amplified with normal PRC.
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Describe the process of PCR:
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1. Denaturation: Heat to near boiling to denaturate DNA
2.Annealing: Cool mixture to 45-65 degrees to allow primers to bind to separated DNA strands 3. Extension: Increase the temperature to 72 degrees to allow the Taq P. to activate and add nucleotides |
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What is Taq P.
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It is a DNA polymerase that is heat stable
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What components are needed for PCR reactions?
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1. Taq P.
2. Deoxyneoclitides 3. Synthetic Oligonucleotides |
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How many cycles of PCR does a template undergo normally?
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20-40 cycles. After 40 cycles, components become depleted.
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What is "hot start" primer and what problem does it solve?
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"Hot Start" primer is primer that is added only after DNA denaturation in order to prevent unwanted amplification of products other than the target DNA
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How do we control mutations resulting from PCR
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Maintain low nucleotide concentrations
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What three forms do correct PRC products take?
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1. Double-stranded with no dA at either end
2. dA at only one end of the DNA strands 3. dA at both ends of the DNA strands |
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How do we prevent the attachment of dA to PCR products?
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Use primers that flank restriction enzymes
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What is CODIS?
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Colective DNA Index System
Began in 1990 by the FBI A catalog of the DNA of all of convicted criminals or suspects in the USA |
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How is PCR used in forensics?
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It is used to amplify polymorphic regions of DNA
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How is AIDS characterized?
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It is characterized by progressive deterioration of a patient's immune system allowing outside agents to invade the body
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