...England Biolabs (NEB) #R0552S EcoRI NEB #R0101S NEB buffer 1 NEB #B7001S NEB buffer 2 NEB #B7002S T4 DNA...DNA ligase NEB #M0202S T4 DNA ligase buffer NEB #B0202S DH5 alpha competent cells Invitrogen #18258-012...concentration, use 1 μL) 2 μL 10x NEB T4 DNA ligase buffer 1 μL NEB T4 DNA ligase add ddH 2 O to bring...catalog # DNA Miniprep Kit Qiagen #27104 EcoRI NEB #R0101S NcoI NEB #R0193S Agarose Sigma #A9539 D.2 Screening...enzyme manufacturer's resources, such as NEB’s online tool NEBcloner (Link opens in a new window) , to help...: 5 μL Forward oligo 5 μL Reverse oligo 5 μL 10x NEB buffer 2 35 μL ddH 2 O Incubate for 4 minutes at ...TRC-cloning vector (maxiprep or miniprep DNA) 5 μL 10x NEB buffer 1 1 μL AgeI add ddH 2 O to bring to 50 μL ...

...restriction enzymes, visit (Link opens in a new window) NEB . Restriction enzyme digestion is commonly used in... in a new window) double digest finder tool from NEB . In a 1.5mL tube combine the following: DNA Restriction... sticky ends. See (Link opens in a new window) NEB's compatible cohesive ends chart . If you cannot find...restriction sites. See (Link opens in a new window) NEB's table of methylation sensitive restriction sites...concentration. Learn more at (Link opens in a new window) NEB's website about star activity . If you are digesting...

...time. Resources and References New England Biolabs (NEB) (Link opens in a new window) sells DNA Assembly ... also sell ET SSB (Link opens in a new window) . NEB has other resources, such as a primer design tool...Assembly kits and master mixes, including NEBuilder HiFi (Link opens in a new window) and Gibson Assembly (Link...

... an endonuclease negative E. coli strain such as NEB stable. Dilute 1:3 (µg DNA:µg PEI) in 500 µL total...

... an endonuclease negative E. coli strain such as NEB stable. Make a mixture of a total of 500 μL PEI-OptiPro...

...a linear piece (see (Link opens in a new window) NEB for more information). Thus, we recommend that you...