TIN: Week 5

Question 1

Plasmids

Answer 1

Small circles of DNA mainly found in bacteria and can replicate independently from the host.
Plasmids in the lab are designed to introduce foreign DNA into another cell (which is called Vector).
Plasmids have promotor
Need antibiotic resistance
May have a eukaryotic resistance marker too
No introns or up/downstream sequences
Can only express one isoform

Question 2

Cloning

Answer 2

take gene out of plasmid and insert it into desired plasmid

Question 3

Promoter

Answer 3

where transcription of a gene is initiated

Question 4

Transcription

Answer 4

when a section of DNA is copied to make RNA. RNA may then be used as a template to make a protein.

Question 5

PCR

Answer 5

Polymerase Chain Reaction

Tool used for copying or amplifying gene in original plasmid

Question 6

Primer annealing

Answer 6

allows primers to bind to start & end of the DNA sequence we want to amplify

Question 7

Primer Extension

Answer 7

Synthesizes a new strand of DNA (this doubles the amount of DNA)

Question 8

What are the steps of PCR?

Answer 8

1) DNA denaturation
2) Primer annealing
3) Primer extension

(heating, cooling, warming DNA)

Question 9

Ligation Reaction

Answer 9

Joining of two DNA fragments though the formation of a phosphodiester bond.
Ligase is an enzyme that repairs single and double strand breaks that occur during DNA replication

Question 10

What is SDS-PAGE used for?

Answer 10

uses electrophoresis to separate proteins by molecular weight
works by applying an electric field across a polyacrylamide gel