Everything lmap

Question 1

What is an enzyme that has a specific DNA sequence(site) when at recognition it cuts the DNA?

Answer 1

Endouclease

Question 2

What is a Ligase(protein-wise), what bonds does it form, and where does it form it

Answer 2

What enzyme forms the phosphodiester bonds between the sugar and phosphate group to form the sugar-phosphate backbone? (Also has the ability to bond two strands of separate DNA together)

Question 3

What endonuclease is used in CRISPR-Cas9 system

Answer 3

Cas9

Question 4

What is a Ligase(protein-wise), what bonds does it form, and where does it form it

Answer 4

What enzyme forms the phosphodiester bonds between the sugar and phosphate group to form the sugar-phosphate backbone? (Also has the ability to bond two strands of separate DNA together)

Question 5

What are the processes in PCR, what are the inputs of each process, and at what temperature are they done in?

Answer 5

Denaturation(93)[The DNA sample], Annealing(53)[primers], Elongation(72)[Free nucleotides, Taq polymerase]

Question 6

What is gRNA used for?

Answer 6

gRNA is used by Taq polymerase to find a sequence that is complementary to the gRNA and adjacent to the PAM sequence.

Question 7

What is the DNA synthesizer in PCR, why is it specifically used and what are its heritable origins?

Answer 7

Taq polymerase, because of its bacterial origins it, has high heat resistance meaning that at high temperatures it does not denature making it very efficient for PCR.

Question 8

What is the importance of the PAM sequence?

Answer 8

The PAM sequence ensures that Cas9 does not cut the bacteria’s own DNA as bacteria DNA does not have the PAM sequence.

Question 9

What is sgRNA and how is it used?

Answer 9

sgRNA is used in similar fashion to gRNA but the main difference is that while gRNA is made naturally, sgRNA is made in a lab

Question 10

Which type of end of DNA(after being cut) is most favorable and why?

Answer 10

Sticky ends because it allows for them to be annealed to complementary bases from other pieces of DNA

Question 11

What are the advantages of Taq polymerase?

Answer 11

Taq polymerase is a kind of DNA polymerase that can withstand high temperatures due to its bacterial origin. The ability of Taq polymerase to withstand high temperatures is why its favourable in PCR

Question 12

What is the role of restriction enzymes(restriction endonuclease)

Answer 12

the protein used to cleave DNA at a specific recognition site.
In order to cut the DNA, restriction enzymes cleave the phosphodiester bond of the sugar-phosphate backbone which holds nucleotides together.

Question 13

protospacer

Answer 13

short sequence of dna extracted by cas1 and cas2 which has yet to be incorporated into the crispr gene

Question 14

CRISPR-Cas9 (definition)

Answer 14

a complex formed between gRNA and Cas9 which can cut a target sequence of DNA. Bacteria use this complex for protection from viruses and scientists have modified it to edit genomes

Question 15

CRISPR Cas9
Step 1 (Exposure)

Answer 15

-bacteriophage injects its DNA into a bacterium, which identifies the viral DNA as a foreign substance.
-cas 1 and cas 2 cut out a short section of the viral dna ~30 nucleotides long. this section of the viral DNA is called the protospacer. this protospacer can be introduced into the crispr gene

Question 16

CRISPR Cas9
Step 2 (Expression)

Answer 16

-gRNA binds to Cas9 to create the CRSIPR-Cas9 complex which is directed to any viral DNA which is complementary to the gRNA.
-the gRNA forms a hairpin loop structure from the transcribed palindromic repeats either side of the repeater.

Question 17

CRISPR Cas9
Step 3 (Extermination)