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What is your favorite method for getting inserts into vectors?


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#1 labtastic

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Posted 22 July 2017 - 01:50 PM

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. :)

#2 pito

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Posted 26 July 2017 - 11:31 AM

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. smile.png

 

Depends on the vector as well.

 

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


If you don't know it, then ask it! Better to ask and look foolish to some than not ask and stay stupid.


#3 labtastic

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Posted 02 August 2017 - 10:51 AM

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



#4 pito

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Posted 03 August 2017 - 03:31 AM

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.


If you don't know it, then ask it! Better to ask and look foolish to some than not ask and stay stupid.





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