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hep-2 & Hep-G2 cell line - difference between hep-2 & Hep-G2 cell line (Dec/15/2004 )

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I have now transfected my HepG2s using the reverse transfection method. I've done some optimisation and had best results (>80% I'm not kidding!) when using 3ug oligonucleotide (oligo is Cy5 labelled) and a 1:2 ratio, leaving the transfection mixture 3h to incubate.

If anyone wants to see the raw data please email me.

LeserattePD

QUOTE (Hooly @ Feb 18 2005, 06:22 PM)
I just read about a technique called 'reverse transfection' for getting siRNA into those difficult-to-transfect cell lines. Basically this means that instead of plating the cells and leaving them alone for 24 hours before transfecting, you resuspend them after trypsinisation in siRNA/liposome soup and *then* plate them.

Haven't tried it, but it was in a newsletter from Ambion - you can find the details at the following link:

http://www.ambion.com/techlib/tn/121/9.html

They specifically say that this is a great way to get really good delivery/silencing in HepG2.

-LeserattePD-

HI,

I just start to work on HepG2 cell transfection with DNA. The reverse transfection you mentioned here is very interesting. Could you please provide more information, such as the transfection reagent, the incubation time,...? Thank you very much for your time and help.

Xin


QUOTE (LeserattePD @ Jun 6 2005, 10:27 AM)
I have now transfected my HepG2s using the reverse transfection method. I've done some optimisation and had best results (>80% I'm not kidding!) when using 3ug oligonucleotide (oligo is Cy5 labelled) and a 1:2 ratio, leaving the transfection mixture 3h to incubate.

If anyone wants to see the raw data please email me.

LeserattePD

QUOTE (Hooly @ Feb 18 2005, 06:22 PM)
I just read about a technique called 'reverse transfection' for getting siRNA into those difficult-to-transfect cell lines. Basically this means that instead of plating the cells and leaving them alone for 24 hours before transfecting, you resuspend them after trypsinisation in siRNA/liposome soup and *then* plate them.

Haven't tried it, but it was in a newsletter from Ambion - you can find the details at the following link:

http://www.ambion.com/techlib/tn/121/9.html

They specifically say that this is a great way to get really good delivery/silencing in HepG2.

-capricy-