Manipulation of DNA - Ligases Flashcards
1
Q
what do ligases do
A
ligases join nucleotides together
2
Q
why are low tempratures used
A
low tempratures ensure stable base pairing
3
Q
what cofactor is used by T4 ligase and what does it require
A
atp and a 5’ phosphate
3
Q
what chemical groups are joined by ligases
A
phosphate and oh
4
Q
are blunt ends or sticky ends better for ligation
A
sticky ends are ligated better
5
Q
what can be used to joing blunt and sticky ends
A
linkers or adaptors are used
6
Q
how do linkers work
A
linkers contain blunt ended fragments with restriction sites, to create and overhang when cut
7
Q
what is topo cloning
A
topo cloning uses topoisomerase to unwind and break sites to inset a genetic sequice where other forms wouldn’t work, such as lacking phosphate groups
