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13 Cards in this Set
- Front
- Back
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Define PCR
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A technique that results in nucleic acid amplification of a specific DNA region(target region)
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Describe the two main types of molecular diagnostic techniques
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PCR-nucleic acid amplification
Probe-nucleic acid hybridization |
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Describe the composition of a nucleotide
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Sugar, phosphate group, base
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Identify the four DNA bases and classify each as either a purine or a pyrimidine
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Adenine and Guanine-purines
Cytosine and Thymine-pyrimidines |
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Identify the required components for PCR
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1. Enzymes
2. dNTP's(deoxynucleoside triphosphates) 3.Primers 4. Buffers and MgCl 5. Mineral Oil/PCR gems 6. Specimens |
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Explain the purpose of the polymerase enzyme
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Taq- can withstand very high heat. Most enzymes would denature in these conditions rendering the test ineffective
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Identify potential sources of contamination during a PCR procedure
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Anything in the environment (even a few bacteria can amplify millions of DNA segments)
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During the denaturation phase of PCR the sample must be heated to this temperature
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95
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During the annealing phase of PCR the sample must be cooled to this temperature to allow for the attatchement of what component?
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55, primers
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During the extension phase of PCR the sample must be heated to this temperature to allow for the addition of what component?
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72, bases added by polymerase
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Identify and breifly explain various detection methods used for identifying PCR
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1. Gel electrophoresis-band migration is unique comparaison with a DNA ladder
2. ELISA-monoclonal antibodies against PCR products tagged with and enzyme 3. Flourescence, Chemiluminescence |
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Identify a common DNA probe
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A probe labeled with a cheminolumiscent compound bound to magnetic particles
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Briefly explain the procedure for the DNA probe technique
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1. Add reagent to digest any unhybridized probe
2. Usa a magnet to pull all hybridized DNA to the side of the tube. 3. Wash sample to remove any digested unhybridized probe 4. What remains will contain only the hybridezed DNA 5. Measured by chemiluminometer |
