Genomic Technologies

Question 1

What are restriction endonucleases?

Answer 1

enzymes that cleave DNA at specific sequences

E.g. EcoRI

Question 2

What is gel electrophoresis?

Answer 2

A way of visualizing DNA that separates fragments based on size

Question 3

What is recombinant DNA?

Answer 3

Any form of linking two initially separate pieces of DNA.

E.g. adding sequences to vectors

Question 4

What is molecular cloning?

Answer 4

A DNA fragment is inserted into a vector that can replicate independently in a host cell

Question 5

What are three kinds of vectors designed to accommodate large DNA inserts?

Answer 5

Cosmids

Bacterial Artificial Chromosomes

Yeast Artificial Chromosomes

Question 6

How is RNA cloned?

Answer 6

By using reverse transcriptase to copy it into DNA. The resulting DNA (cDNA) is ligated to a vector DNA

Question 7

What is Whole Exome Sequencing?

Answer 7

DNA sequencing of all of the coding exons in the genome

Question 8

What is RNA sequencing?

Answer 8

Characterizing the transcriptome by sequencing mRNAs.

Question 9

What is Chip Sequencing?

Answer 9

Sequencing the binding sites for different transcription factors (DNA protein interaction)

Question 10

What is Genomic medicine?

Answer 10

Using a patients DNA sequence for monitoring and guiding therapy

Tumor therapy

Question 11

How can sequencing be utilized with epigenetics?

Answer 11

Assessing the epigenome using bisulfite sequencing.

Allows detection of fetal syndromes

Question 12

What is metagenomics?

Answer 12

The assessment of the microbiome using DNA sequencing

Question 13

What is the significance of the 16s rRNA gene in metagenomics?

Answer 13

The 16s rRNA is highly conserved, but has variable regions that are species specific. Allows study of evolutionary relationships and ID

Question 14

What is nucleic acid hybridization?

Answer 14

DNA strands are initially separated by high temperature, and then re-form helices when cooled

Question 15

What is Southern blotting?

Answer 15

Cloned DNA is labeled with radioactive NTs or fluorophores.

Labeled DNA is then used as a probe that hybridizes with complement DNA or RNA

Question 16

What are DNA Arrays?

Answer 16

DNA “microchips” used to detect gene expression levels or SNP detection

Question 17

What is PCR?

Answer 17

An in vitro method for the enzymatic synthesis of specific DNA sequences

Question 18

What three steps are repeated in PCR?

Answer 18

Denaturation

Annealing

Extension

Question 19

What is Real-time PCR or Quantitative PCR?

Answer 19

The formation of PCR product is monitored continuously by means of fluorescent primers

Utilized in mRNA analysis for gene expression

Question 20

What are four applications for PCR?

Answer 20

• DNA amplification
• Assessing Gene expression
• Genotyping
• in situ PCR

Question 21

What is in situ PCR?

Answer 21

The amplification of the target nucleic acid sequences in cells or tissue samples that are fixed and permeabilized.

Assesses if and where a gene is expressed

Question 22

How can gene function be studied in eukaryotes?

Answer 22

By introducing cloned genes into the germ line of multicellular organisms.

E.g. Transgenic mice

Question 23

What is gene knockout?

Answer 23

Eliminating the activity of the normal gene copy to determine its role.

Question 24

What is in vitro mutagenesis?

Answer 24

Inactivating certain genes in mouse ESCs, which grow into mice where the effect of the inactivation can be studied.

Question 25

What is the Cre-Lox system?

Answer 25

Takes advantage of a site=specific recombinase to introduce gene deletions, inversions or translocations.

Allows for conditional mutagenesis

Question 26

What is the CRISPR/Cas

Answer 26

Allows for targeted gene editing.

Question 27

What are MicroRNA?

Answer 27

Highly conserved mechanism that targets mRNAs in the 3’ UTR to regulated protein expression post-transcriptionally

Question 28

What are small interfering RNAs and how do they work?

Answer 28

Inhibits mRNA by activating endonuclease that digests a dsRNA target containing mRNA