Tips on ethanol precipitation of nucleic acids and wash - (Mar/13/2005 )

I'd just add EtOH as if it were a pure precipitation starting with a solution without any EtOH in there. I also don't think that isoprop would interfere too much with EtOH, but that's just a feeling. Both compounds precipitate the DNA by the same mechanism (after all, they're both alcohols, so from chemical perspective not TOO different).

For the amount of glycogen to add to a precipitation: the final concentration should be anywhere between 0,05 and 1 µg:µl (according to Fermentas...) Pretty wide range, but you can easily see minute amount of glycogen after precipitation. Indeed, add this before isoprop/EtOH.

What about using Glycogen and RNA from adipocytes?

And what are the qualitative differences between sodium acetate and glycogen?

Hi all

to get rid the ethanol, I usually cover up the top of microcentrifuge with parafilm and then make several small holes by using tips..after that place it into vacume centrifuge to enable the ethanol to evaporate more quickly

That's what I thought too, but my boss did not believe me

Hi Bioforumers,

I have to sequence small human DNA (600 bp). I am using the Beckman Coulter Dye Terminator Cycle Sequencing with Quick Start Kit.

After the DNA sequencing reaction, I have to precipitate the DNA. So I will use Sodium Acetate (3M), EDTA (100mM) and glycogen (20 mg/ml). The problem is that I have not enough glycogen, so I want to buy it.

You can find a lot of different glycogen on the manufacturers' catalogs, in solid powder or in aqueous solution, and from different species (Oyster, Mussels, Bovine, Rabbit, Slipper limpet). The powder costs 50€ for 5 g, but the aqueous solution (35 mg/ml) costs 59€ for 0.5 mL !! This aqueous solution is said DNASE, RNASE, nuclease free, but it's 337 fold more expensive...

There is also different types of glycogen (type II, III, VII, VIII, IX, XI). Does anyone knows the differences between them?

Can you help me to find the good product asap?
I have already begun my sequencing reactions...

Thanks a lot
SB

Hi Bioforumers,

I have to sequence small human DNA (600 bp). I am using the Beckman Coulter Dye Terminator Cycle Sequencing with Quick Start Kit.

After the DNA sequencing reaction, I have to precipitate the DNA. So I will use Sodium Acetate (3M), EDTA (100mM) and glycogen (20 mg/ml). The problem is that I have not enough glycogen, so I want to buy it.

You can find a lot of different glycogen on the manufacturers' catalogs, in solid powder or in aqueous solution, and from different species (Oyster, Mussels, Bovine, Rabbit, Slipper limpet). The powder costs 50€ for 5 g, but the aqueous solution (35 mg/ml) costs 59€ for 0.5 mL !! This aqueous solution is said DNASE, RNASE, nuclease free, but it's 337 fold more expensive...

There is also different types of glycogen (type II, III, VII, VIII, IX, XI). Does anyone knows the differences between them?

Can you help me to find the good product asap?
I have already begun my sequencing reactions...

Thanks a lot
SB