Site directed mutagenesis

WS via (by novalidaddress from
Mon Aug 15 03:50:43 EST 2011

you will bite yourself in the a** when you later discover that there
was a mutation somewhere else in your sequence. Considering the cost
of commercial sequencing (the cheapest way are those "packages" where
they just run the reaction and you get the raw data and do the
evaluation by yourself, there is free software available like "finch
tv"), sequencing your constructs is a must before you derive any data
from them that you want to publish/patent or which are important in
any other context.

I had learned my lesson once by finding out almost too late that one
of the primers I had used was missing a base (still in the time of
running your own DNA synthesizer and sequencing with gels and 35S)

my 2c

On 15 Aug., 04:05, mnr mnr <mnr... from> wrote:
> If I were to go down the Phusion road, will I need to sequence the entire of
> my gene (4.5 kbp) or just the mutated region to make sure the protein
> finally has mutated in the right places?

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