week 2

Question 1

bystander edits

Answer 1

when off target sequences (sequences similar to the sequence we want) also gets cut and is near target sequence

Question 2

deaminase

Answer 2

enzyme that converts AT base pair to CG base pair. used in one of the uses of crispr that leads to site-specific modifications

Question 3

plasmid dna

Answer 3

codes for grna and cas9

Question 4

3 methods to transfect gRNA and cas9 into s2 cell

Answer 4

1. use plasmid containing gRNA and cas9 dna
2. use gRNA and cas9 rna
3. use gRNA and cas9 protein

Question 5

how to inject gRNA and cas9 into s2 cell

Answer 5

physically (like injecting), using a virus, or using synthetic material

Question 6

primary cell

Answer 6

cell isolated from plant or animal tissue

Question 7

primary cell must undergo … to become a continuous cell line

Answer 7

crisis (telomerase activation)

Question 8

eagle’s media is used for… and contains

Answer 8

growing cells. however, we’re using schneider’s insect media to grow s2 cells
amino acids, vitamins, glucose, salts, modified eagles medium

Question 9

continuous vs transformed cell line

Answer 9

continuous: spontaneous transformation
transformed: comes from tumor

Question 10

complete media needs…

Answer 10

serum (has growth factors, nutrients, and 5 or 10% horse, calf, or fetal calf serum)

Question 11

types of transfection (transfer dna/rna into cells)

Answer 11

transient and stable

Question 12

what will PCR do for us in this exp

Answer 12

amplify genomic DNA so that we can check if smo gene has indels

Question 13

the 3 steps of PCR and their corresponding temps

Answer 13

1. denaturation- heating up dna so that H bonds between the starnds can break. done at 98C
2. annealing: primers base pair to the now separated strands. done between 58-65C
3. extension: taq polymerase extends primer to form new dna strand. done at 72C