adding glycogen to isoprop for precipitation? - (Aug/10/2006 )
Hello all,
I have a quick question, I have A LOT of precipitations to do in the near future (more than 500), and adding glycogen to the precipitation mixture increases DNA and RNA yield (I have to do both, and samples contain very small amounts of nucleic acids, experiments have to do with determining detection limits). But, as adding glycogen to the reactions themselves would mean roughly 500 extra pipettings, I was wondering if it would harm my glycogen (or the precipitation procedure as a whole) if I would add it to my isopropanol in advance... It would save me a lot of time, but if it would make my precipitation perform less, I would not do it of course.
Any thoughts?
Thanx!
I have a quick question, I have A LOT of precipitations to do in the near future (more than 500), and adding glycogen to the precipitation mixture increases DNA and RNA yield (I have to do both, and samples contain very small amounts of nucleic acids, experiments have to do with determining detection limits). But, as adding glycogen to the reactions themselves would mean roughly 500 extra pipettings, I was wondering if it would harm my glycogen (or the precipitation procedure as a whole) if I would add it to my isopropanol in advance... It would save me a lot of time, but if it would make my precipitation perform less, I would not do it of course.
Any thoughts?
Thanx!
If nobody knows (at least I have no idea, but it sounds interesting) it would be a nice little experiment. I'm interested in the results, if you do it
. May be Free_33 could know, if not… 
I think you should do a little test as jobglobin suggested, do two assays with the same sample, one of them adding glucogen to isopropanol and the other with the normal procedure.
Good look!
I'm not a 100% sure but I think the glycogen might precipitate in the presence of isopropanol.
You could try adding a couple of ul glycogen to a small volume of isoprop, mix, and spin it down to see if you have a pellet. If you do, you know you've lost your short cut.
Yes they are right, you could try a little pilot experiment.
hi
glycogen is supposed to be added with the nucleic acids, vortexed with, and then adding the isopropanol.
I don't know about glycogen precipitation and thus can not says much on that.
What i can suggest you, is using a repetitor. I mean we have a pipett in the lab by which you pipett once and then release 50µl as many as you want. So i think about preparing a mix water and glycogen, and add 50µl to your RNA preparation. Then proceed with isopropanol addition.
If you don't have automatic repetitor, i think you can do that manually (seems equivalent to adding a 30µl PCR mix or sthg like that.
You could try adding a couple of ul glycogen to a small volume of isoprop, mix, and spin it down to see if you have a pellet. If you do, you know you've lost your short cut.
My curiosity got the better of me. I've just added 2ul gycogen (40ug) to 100ul isopropanol and with some gentle pipetting it formed a huge white precipitate before my eyes. The glycogen has to be mixed with the DNA before the ETOH/isoprop. precipitation.
i wish to do that, but didn't took the time. Thanks for the information
i wish to do that, but didn't took the time. Thanks for the information
Thanks Fred_33...you're too busy solving everyone's problems to have the time!
now you know how glycogen helps to precipitate DNA 