发信人: adeno (天夺其魄), 信区: Biology
标 题: Re: CaP transfection
发信站: Unknown Space - 未名空间 (Sat Feb 7 20:24:48 2004) WWW-POST

It's not availbale in the web, but in the book "current protocols in molecular
biology". Most molecular biology labs should have it. I am sure you can find
one in either your lab or a nearby one. They are 3 huge red binders. A concise
version "short protocols in molecular biology" also exist.

Basically the method is the same as HEPES one. If you cannot find it I can
find you a recipe when I go back to lab.

As for toxicity, Roche's fugene 6 is famous for lower toxicity than other
liposomal method, which you may want to give a try.

【 在 raison (raison) 的大作中提到: 】
: thanks a lot adeno. i'll try BES. can you give the link of the protocol in
: current protocols in molecular biology?
:
: as for cell type - dependent toxicity, you are right. i'm using cortical
: neurons isolated from embryonic mice, which is well-known for susceptibility
: to transfection
:
:
:
: 【 在 adeno (天夺其魄) 的大作中提到: 】
: : Try that alternative protocol in current protocols in molecular biology
: which
: : use BES instead of HEPES.I got much higher efficiency than with HEPES,
and
: I
: : didn't find any obvious toxicity.
: :
: : Toxicity may also be cell type specific. Regular HEK293 and Hela cell
which
: I
: : used seem all right with CaP
: :
: : 【 在 raison (raison) 的大作中提到: 】
: : : anyone done CaP transfection successfullly?
: : :
: : :
: : : i tried according to this protocol
: : :
: : :
: http://gweb1.ucsf.edu/labs/finkbeiner/Protocol/protocol_list/Ca_P_TX_R.shtml
: : :
: : : the transfection efficiency is much higher than expected, but the
toxicity
: : is
: : : unpleasantly high as well. around 1/3 of cell death.
: : :
: : : do you guys have any suggestions? thanks
: : :
: : :
: :
:
:

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