Protocols

[Alisa]

Site-directed mutagenesis:
I'm pretty sure the kit I've previously used is the Stratagene
QuikChange kit. Results varied. When it did work, it worked well. The
manual can be found at:
http://www.stratagene.com/manuals/200518.pdf

We can get a kit with 10 reactions (which includes 5 control
reactions) or 30 reactions.
There are other protocols, including ones that are similar to
QuickChange, but without the kit. There's a guy in my lab that seems
to know a lot about this, so I'll ask him and get back to you. If we
do decide to get this kit, most of the stuff we need will be provided.
We will, however, need to get our own
-well designed primers
-PCR tubes
-PCR machine
-media and equipment (flame, spreader, eppies) for transformations
-heating block.
Oh, and X-gal and IPTG are recommended. I've never used them though. I
don't think we even use the bacteria they provide us. We will also
need -20 storage. I don't know that we'll be able to get one for our
exclusive use though.
There's probably more - I'll add them when I remember.

[Craig]

I uploaded several protocols and how to make up some solutions which I
thought might come in handy. Some of the stuff we probably will not
use but just in case. Except for the PCR protocol and the extraction
method which I wrote up, the source is as follows:

T. Maniatis et al. Molecular Cloning A Laboratory Manual. First
Eddition.

I forgot to write up what materials are needed for each reaction, and
I will do that next. I will leave out standard things probably found
in a lab e.g. autoclave, bunsen burner, etc.