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Amplification Techiques

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wayla's version from 2016-11-16 23:58

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Question Answer
Amplification of specific, small DNA sequences using a primer extension of complimentary strands of DNA is what test?Polymerase Chain Reaction (PCR)
What are the advantages of doing PCR testing?It is a fast test (2-5 hours), DNA or RNA can be amplified, and it is automatized and standardized.
How many steps and cycles are in the PCR test?There are 3 steps which are repeated 20-30 times
Describe what occurs reverse transcription PCR.Single-stranded RNA template is converted to a complementary DNA molecule by RNA dependent DNA polymerase.
When is reverse transcription PCR useful?For determining viral load of RNA viruses.
Describe what occurs in multiplex PCR.Amplification of multiple targets.
What is an advantage of multiplex PCR?It is cost effective and conserves time and effort in the lab.
What does a negative control for PCR consist of?Contains all reagents and a DNA template lacking the target sequence
What does a positive control for PCR consist of?Contains all reagents and a known target-containing DNA template
What is the purpose of an internal control for PCR ?Ensures reaction mixture is working and that there are no inhibitors.
What are the components of PCR?-A target (template) DNA (ssDNA) or RNA in sample -Primers
-DNA polymerase
-Deoxynucleotide triphosphates (dNTPs)
What are some disadvantages to PCR testing?-Must know the sequence of the DNA of interest
-Highly susceptible to contamination or false amplification
-Amplification may not be 100% specific
-Analysis and product detection usually takes longer than the PCR reaction itself
Which PCR amplifies specific sequences of DNA from a large complex mixture of DNA?Nested PCR
Which PCR uses SYBR-green fluorescent dye?Real-time/Quantitative PCR
What are the 2 types of probe amplification methods?Ligase chain reaction and strand-displacement amplification
What lessens the risk of target contamination in the branched chain DNA detection method?The number of target sequences does not change and is bound to a large amount of signal.
What kind of polymerase does PCR use?Thermostable DNA polymerase
As copy number increased, what happens to the signal?It increases
What does the term isothermal mean?Requires one temperature
In branched chain DNA detection, how often does the number of target sequences change?It stays the same.
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