PCR_and_Sequencing_Flashcards
What factors influence the choice of DNA extraction technique?
Target nucleic acid, starting material, expected results, and downstream applications.
What are examples of target nucleic acids?
RNA, circulating DNA, or other nucleic acids.
How does the starting material affect DNA extraction?
Tissue, cell culture, or biopsy dictates the approach to release nucleic acids.
Why is the extraction method important for downstream applications?
It must be compatible with the intended molecular biology test.
What is PCR used for?
Amplifying a specific DNA region in vitro to obtain sufficient quantities for study.
What is the outcome of PCR amplification?
Exponential replication of a DNA template.
What are the main steps of PCR?
Denaturation, hybridization, and polymerization.
What happens during denaturation in PCR?
Double-stranded DNA is separated into single strands by heating.
What occurs during hybridization?
Primers bind to the single-stranded DNA, defining the region to amplify.
What is polymerization in PCR?
DNA polymerase extends primers, creating complementary strands.
What are primers?
Short DNA sequences (forward and reverse) that bind to the target DNA region.
What is a thermocycler?
A machine that controls temperature cycles for denaturation, annealing, and extension.
What are PCR tubes made of?
High-quality polypropylene for efficient heat transfer.
What is the purpose of a PCR buffer?
To provide optimal conditions for the reaction.
How is electrophoresis used in PCR?
To separate DNA fragments by size using an electric charge.
