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Quiz 10

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Justine C.
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The first step of PCR requires that the DNA to be amplified is

denatured.

The chain-termination method of sequencing relies on the formation of partial copies of the DNA fragment that terminate at each position in the sequence. The fragments are separated by size and the terminating nucleotide is identified by

the position in the capillary and a unique fluorescent tag on each nucleotide.

Reverse genetics is defined as

the inactivation of a gene of interest to determine its function.

A genomic library is a collection of plasmids, each containing different chromosomal fragments from the genome of an organism. A cDNA library, on the other hand, contains only the ___________ from the genome of an organism.

coding sequences of genes

Overexpression of genes in zebrafish can be achieved by______.

injection of plasmid DNA into zebrafish embryos.

If a protein with a specific function is isolated, how can the encoding gene be identified?

sequencing

DNA migrates through the pores of an agarose gel toward the ___________ electrode; larger fragments travel more ___________.

positive; slowly

Unlike RNA-seq, in situ hybridization can reveal what about gene expression?

temporospatial

Restriction enzymes cut DNA

at a specific sequence, whether in cells or in a test tube.

DNA fingerprinting uses DNA sequences in the genomes of individuals that are hypervariable, and the figure below depicts the results of fingerprinting four individuals. The regions analyzed are called

short tandem repeats.

Study Notes

PCR and DNA Sequencing

  • The first step of PCR requires that the DNA to be amplified is prepared.
  • The chain-termination method of sequencing relies on the formation of partial copies of the DNA fragment that terminate at each position in the sequence.
  • The terminating nucleotide is identified by separating the fragments by size.

DNA Libraries

  • A genomic library is a collection of plasmids, each containing different chromosomal fragments from the genome of an organism.
  • A cDNA library contains only the expressed genes from the genome of an organism.

Gene Expression and Regulation

  • Overexpression of genes in zebrafish can be achieved by microinjection of mRNA.
  • If a protein with a specific function is isolated, the encoding gene can be identified by using the protein to screen a library.

Agarose Gel Electrophoresis

  • DNA migrates through the pores of an agarose gel toward the positive electrode.
  • Larger fragments travel more slowly.

Gene Expression Analysis

  • Unlike RNA-seq, in situ hybridization can reveal the spatial distribution of gene expression.

Restriction Enzymes and DNA Fingerprinting

  • Restriction enzymes cut DNA at specific recognition sites.
  • DNA fingerprinting uses DNA sequences in the genomes of individuals that are hypervariable.
  • The regions analyzed in DNA fingerprinting are called variable number tandem repeats (VNTRs).

Explore the basics of PCR, DNA sequencing, and DNA libraries. Learn about the preparation of DNA, chain-termination method, and genomic libraries.

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