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Genetics · Genetics

Genetics: Genomic Technologies and Sequencing

PCR, next-generation sequencing, and CRISPR gene editing technology concepts at the introductory genomics level.

30 cards · basic cards · AI-written, checked twice. Edit anything.

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What does PCR stand for?
Polymerase Chain Reaction
What is the main purpose of PCR?
To amplify a specific segment of DNA into millions of copies
Who invented PCR in 1983?
Kary Mullis
Which enzyme is used to synthesize new DNA strands during PCR?
Taq DNA polymerase
From what organism is Taq polymerase originally derived?
Thermus aquaticus, a heat loving bacterium
What are the three steps of a single PCR cycle, in order?
Denaturation, annealing, extension
What happens during the denaturation step of PCR?
The DNA is heated (about 95C) so the double strand separates into two single strands
What happens during the annealing step of PCR?
Primers bind to their complementary sequences on the single stranded DNA template
What happens during the extension step of PCR?
DNA polymerase adds nucleotides to the primer, synthesizing a new complementary DNA strand
What is a primer in PCR?
A short, synthetic single strand of DNA complementary to the start of the target sequence, needed to begin DNA synthesis
What does NGS stand for?
Next-Generation Sequencing
What is the key advantage of NGS over Sanger sequencing?
It sequences millions of DNA fragments in parallel, rather than one at a time
What method does Sanger sequencing use to stop DNA synthesis at each base?
Chain termination using dideoxynucleotides (ddNTPs)
What is the basic principle behind Illumina sequencing?
Sequencing by synthesis: a fluorescent signal is detected each time a base is added to a growing DNA strand
In sequencing, what is a 'read'?
A short stretch of DNA sequence produced by a sequencer

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