biotech

Question 1

Define Probe

Answer 1

Single Stranded DNA/RNA that have been manmade to locate genes. Are either fluorescent or radioactive. Is complementary to GOI

Question 2

Define Restriction Enzyme

Answer 2

Enzymes produced by bacteria which cut DNA at specific sites. Used to remove and/or isolate genes.

Question 3

Describe PCR steps

Answer 3

1. Template DNA is heated to 95 degrees to break hydrogen bonds and expose bases.
2. Mixture is cooled to 65 degrees to allow primers to bind to exposed bases.
3. Temperature is raised to 75 degrees to allow nucleotides to bind to exposed DNA bases. TAQ polymerase will join them together. 2x DNA has been made.
4. Repeat process, amplify DNA.

Question 4

Define TAQ DNA polymerase

Answer 4

enzyme from bacteria that can tolerate higher temperature. Used to bind DNA.

Question 5

Define Primers

Answer 5

Short section of DNA that bind to either side of the template strand to prevent double stranded DNA from forming and acting as a binding site for TAQ polymerase.

Question 6

Describe how the GOI is extracted

Answer 6

1. Restriction enzyme cuts DNA at specific location.
2. Gene probe complementary to the GOI is added.
3. They can now be identified and separated (do not need to know how).

Question 7

Define a terminator base

Answer 7

Human edited nucleotide which is flureoscently tagged (each base a different colour), and ‘terminates’ the strand (will always be the final base).

Question 8

Describe Gel Electrophoresis

Answer 8

1. DNA fragments are cut using restriction enzymes.
2. DNA is added to wells of agar gel plate, electrically charged.
   DNA is negatively charged
3. If an electric current is run through the liquid DNA
   is attracted to the positive end
    Smaller DNA fragments move faster
   each person’s DNA gets cut into
   different size fragments which is unique to that
   person

Question 9

Describe DNA sequencing

Answer 9

1. DNA samples are tagged with terminator bases.
2. DNA is run through capillary electrophoresis
3. DNA strands are in size order, the final terminator bases colour tagged, allowing the full strand to be sequenced.

Question 10

Define Short Tandem Repeat (STR)

Answer 10

Non-coding regions of DNA contain short repeated sequences of DNA code, unique to the original. Although similar STR’s indicate genetic relation.

Question 11

Define Genetic Engineering

Answer 11

The techniques and processes used to directly manipulate the genomics of organisms

Question 12

Define Vector

Answer 12

The vehicle used to transport GOI to cell.

Question 13

Define Microinjection

Answer 13

The use of a tiny needle to
directly inject multiple copies of the GOI into the cell

Question 14

Define Electroporation

Answer 14

By shocking the cell the
membrane opens and GOI or plasmids can enter the cell.

Question 15

Define Plasmid

Answer 15

A plasmid is a small, circular, double-stranded DNA molecule that exists separately from the main bacterial chromosome

Question 16

Define AAV

Answer 16

A plasmid is edited to contain the GOI. Harmful Virus DNA is removed, and the plasmid is inserted.
Virus injects their own DNA into human cells.

Question 17

Define Cas-9 enzyme

Answer 17

Cas-9 will pick up pieces of RNA and use it as a guide. Cas-9 reads DNA, if it finds a strand complementary to the guide RNA, it will make a blunt cut on both sides.

Question 18

Explain CRISPR

Answer 18

1. Guide RNA is complementary to sequence needing to be cut
2. Cas-9 enzyme locates sequence needing to be cut, and creates a blunt cut on both ends.
3. Gene is either deleted, or replaced with GOI (added through vector).

Question 19

define transgenesis

Answer 19

transferring genes between species