How much?! - (Sep/04/2006 )

Hi there

Got a bit of a problem, back to the lentiviral work, all protocols say I need 10 microgram of the required siRNA sequence. Now my stock solution is only 1.1 microgram/microlitre. How is it possible to get this amount of siRNA? I'm very new to this this whole molecular biology thing, as I'm an electrophysiologist!

Please help anyone!!!

-wildcat-

9ul x1.1 ug/ul=10 ug

no knowledge of electrophysiology involved. :-)

-genehunter-1-

QUOTE (genehunter-1 @ Sep 4 2006, 04:29 AM)

9ul x1.1 ug/ul=10 ug

no knowledge of electrophysiology involved. :-)

Genehunter, I think 1ug/ul diluted to 10ul gives 1ug in 10 ul not 10ug...

Good joke

-smartsunny-

QUOTE (wildcat @ Sep 4 2006, 02:57 AM)

wildcat you will need to prepare fresh concentrated stock.... How did you prepare your siRNA? Did you prepare dsRNA by in vitro transcrioption and then Dicer it??

-smartsunny-

As far as I can follow what's going on, needing 10 micrograms in total requieres 9 µl of a 1.1 microgram/µl solution.

If you need a solution of 10 µg/µl you want to freshly prepare or increase concentration by precipitation or vacuüm/evaporation.

-vairus-

I agree with others that one needs 9ul of 1.1ug/ul if u need 10ug of siRNA.

But how is this related to lentivirus ? do u have shRNA cloned into lenti vector?

-scolix-

I've got a shiRNA cloned into the lentivirus. It sounds sensible to use the 9microliters/ml instead of making a new stock, as I received this as a gift and wouldn't know where to start either.

Thank you for all your help

T

QUOTE (scolix @ Sep 4 2006, 04:29 PM)

I agree with others that one needs 9ul of 1.1ug/ul if u need 10ug of siRNA.

But how is this related to lentivirus ? do u have shRNA cloned into lenti vector?

-wildcat-

BTW who suggested to use 10ug for knocking down. I havent heard of any such thing as using a certain amount of vector for knockdown.

take care of how u plan ur experiments and also abt the vector (as u suggested its a gift) so dont use it all up.

-scolix-