4 replies to this topic

[sowmi]

Hi all,

I m working on insects.. would like to know if there is an easy and cheap way (No liquid nitrogen.. No RNA later... )  for storage of insect larvae before RNA isolation.

Sample collection takes some time and I would prefer a simple receipe for a storage liquid.

It should keep my RNA safe for atleast 2 hours and should give options for low temperature storage after that..

any suggestions???????????

[AquaPlasmid]

Sowmi,

Would you like to try AquaPreserve?

The basic protocol is: Put larva in AquaPreserve - transport/store at room temp (or freeze down) - homogenize - spin - supernatant to isopropanol - spin to pellet DNA/RNA - DNase I to get RNA.

Cheers,
aquaplasmid

"Finally, total RNA can be extracted from frozen whole blood!"

[sowmi]

hi

thanks for your suggestion.. but i m looking for something much simpler.. (to try till I get the commercial storage solution...)
something conventional..
how did RNA enthusiast survive before all these commercial products?
definitely there should have been some way out.


does ethanol (75-100%) @ 4C help for short term storage? say 1 hour...


I am looking for more replies.. am I a lone crusader here?????????



Cheers..

sowmi..

(Fighting RNase!!!!!!!)

[Mullet]

No, you´re not alone!
we also work with insects, but we use Trizol for storage of samples - snap freeze in liquid N2, then on ice (if in the field) or -80
not very innovative i´m afraid, but easy enough.

sowmi, on Jun 15 2010, 07:08 AM, said:

hi

thanks for your suggestion.. but i m looking for something much simpler.. (to try till I get the commercial storage solution...)
something conventional..
how did RNA enthusiast survive before all these commercial products?
definitely there should have been some way out.


does ethanol (75-100%) @ 4C help for short term storage? say 1 hour...


I am looking for more replies.. am I a lone crusader here?????????



Cheers..

sowmi..

(Fighting RNase!!!!!!!)

[sowmi]

Thanks mullet.. that was reassuring..

But my larvae are too small.. (< 1 mm).. I need to collect almost a hundred for one experiment..
I cant imagine snap freezing each larvae and then placing into ice..     

I need some liquid to hold the larvae for some time before I can freeze them or put away in -80..

Can I have them in trizol for an hour before snap freezing??? will that work????

I know its too unnerving...

Help me out guys..... i ll be eternally indebted......


sowmi...

Looking for replies.. and some reassurance of course!!!!!!!!!!!