DNA synthesis/ synthetic 2

Question 1

what are the 4 major steps of the phosphoramidite oligodeoxynucleotide synthesis cycle

Answer 1

deprotection - removal of DMT allows base addition
base coupling - phosphoramidite added to unprotected 5’ OH
capping - optional
oxidation - oxidation of phosphate triester to phosphate to stabilise

Question 2

what 3 main steps does agilent technologies via inkjet printing follow

Answer 2

deprotection
coupling
oxidation

Question 3

what is sanger sequencing

Answer 3

determining DNA sequences rapidly by primed syntheses with DNA polymerase
selective incorporation of chain-terminating dideoxynucleotides by DNA polymerase

Question 4

what is gibson isothermal assembly and what molecules does it require

Answer 4

assembly of multiple DNA fragments

T5 exonuclease, tag ligase phusion polymerase

Question 5

name 2 artificial restriction enzymes

Answer 5

ZFN - zinc finger nuclease

TALEN

Question 6

what is MAGE and what does it do

Answer 6

Multiplex Automated Genomic engineering

can do 50 different genome mutations at a time for accelerated directed evolution

Question 7

what dies solid phase synthesis mean

Answer 7

molecules are bound on a bead and synthesised step-by-step

Question 8

3 applications for genetic engineering

Answer 8

microbial cell factories - fuels
medicinal/health - tissue engineering
sustainable world - bioremediation

Question 9

3 challenges

Answer 9

escape from lab
bio security and bio hacking
playing god/ devaluing life