Exo' III deletions for sequencing

[Thon de Boer]

Clinton Grant writes:
> 
> 
> 	I am interested to hear from people who have produced Exo' III deletions
> for sequencing: what are the best protocols; are commercial kits the way to go; 
> what are the quirks with the system?
> 	I wish to sewquence a 3kb insert in pBLUESCRIPT KS-.
> 
We are using the Erase-A-Base kit from Promega and it gives variable results.
We had to adjust the ligation step of the protocol (O/N plus extra Ligase,
instead of 1 hour) and one time point can result in clones with random
deletions. But I prefer this over subcloning anytime.

Sincerely, Thon de Boer

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