What is your favorite method for getting inserts into vectors? - (Jul/22/2017 )

What is your favorite method for getting inserts into vectors?

Consider time, cost, efficiency, accuracy and compatibility associated with each method. Does your answer change when considering more complex cloning problems like fusion of multiple dna inserts or making multiple mutations at once?

Examples include (but are not limited to): restriction digest+ligation, in-fusion, Gibson assembly, SLiCE, CPEC, SLIC, gateway cloning, recombination, etc.

I have my own opinions but am curious to hear others' first. :)

labtastic on Sat Jul 22 21:50:57 2017 said:

What is your favorite method for getting inserts into vectors?

Consider time, cost, efficiency, accuracy and compatibility associated with each method. Does your answer change when considering more complex cloning problems like fusion of multiple dna inserts or making multiple mutations at once?

Examples include (but are not limited to): restriction digest+ligation, in-fusion, Gibson assembly, SLiCE, CPEC, SLIC, gateway cloning, recombination, etc.

I have my own opinions but am curious to hear others' first.

Depends on the vector as well.

For me, if possible: restriction digest and ligation. Works all the time.

I agree ligation works well. But it is tedious....

labtastic on Wed Aug 2 18:51:00 2017 said:

I agree ligation works well. But it is tedious....

It all depends on what you want.

If it is a high throughput kind of system and you want to insert lots of genes, then yes, you need something like gateway.