Cloning
Type
Guide
...molecular biology enzymes: 5' exonuclease, polymerase and ligase. 5' exonuclease digests the 5' end of dsDNA...) relies on the 3'-5' exonuclease activity of T4 DNA polymerase. An exonuclease is an enzyme which removes... thus does not need restriction enzymes or an exogenous ligase providing an incredibly quick and easy ...together in one isothermal reaction. First the 5’ exonuclease chews back the 5’ ends of your DNA fragments ...polymerase then closes the gap created by the 5’ exonuclease and finally ligase seals the nicks in the DNA...potential to be completely chewed back by the 5' exonuclease. Gibson Assembly (Image from Plasmid 101: Gibson... DNA strand. In LIC, the T4 DNA polymerase’s exonuclease activity creates “chewed-back” overhangs of 10...