quiz 1 review ( nucleic acids)
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What are nucleic acids?
DNA and RNA
What are the building blocks of both DNA and RNA?
Nucleotides
What are the components of a nucleotide?
Pentose sugar Nitrogenous base Phosphate group
What nitrogen bases are found in DNA?
Adenine Guanine Cytosine Thymine
What nitrogen bases are found in RNA?
Adenine Guanine Cytosine Uracil
What are the basic steps in most molecular assays?
Extraction and purification of nucleic acid Amplification of the target nucleic acid Detection of the amplified target
How are cloned DNA probes made?
Probe is inserted into plasmid vector and placed into bacterium which it replicates.
What can probes be labeled with?
Radioactive element Biotin Enzyme
High stringency occurs under which conditions?
Low salt concentration High temperature Presence of a denaturant (formamide)
What is extracted in the Southern Blot method?
DNA
How are DNA fragments separated in the Southern Blot method?
Gel electrophoresis
Which method is performed on cells, tissues or chromosomes fixed to a microscope slide?
In situ hybridization
Which method evaluates differences in DNA sequences by identifying different lengths of DNA fragments?
Restriction Fragment Length Polymorphism (RFLP)
What is the most common method that uses target amplification?
PCR
What are the 3 steps of each PCR cycle?
Denaturation Primer annealing Primer extension
What is the end product of PCR?
Amplicon
How are the end products of PCR analyzed
Gel electrophoresis
What can cause false-positive results in PCR?
Contamination from carryover or the environment
How can false positives in PCR be prevented
Have different rooms or benchtops for each process Use aerosol-resistant pipette tips
What is the name of the enzyme that extends primers on each template strand in the PCR cycle?
Taq polymerase
What characteristic does this taq polymerase have?
Thermostable
Which molecular technique uses signal amplification?
Branched DNA assay
Which molecular technique uses probe amplification?
Ligand chain reaction (LCR)