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pellet became insoluble after boiling.


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#1 iamdhan

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Posted 31 August 2009 - 01:42 AM

Hello to all!

A colleague in my lab. boiled pellets(that she infected virus) with SDS in 100'C for 5 mins.

The thing is that she didn't mix the pellet with SDS by pipeting before she boiled it.

And to my surprise, it turned out to be a great mistake as the pellet somehow got in the shape and never is soluble at any rate; she tried everything to "destroy" the pellet like pipeting, vortexing, and flicking.

I don't know the full procedure of her experiment, so I might miss some obvious missing step, but nothing seems that wrong to deserve this unchangeable condition of pellet.

What do you think? How come this happens? Could possibly she make it soluble again?

Sorry for lack of detail and also for my broken english. :P


Thx!!

#2 pito

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Posted 31 August 2009 - 05:12 AM

Hello to all!

A colleague in my lab. boiled pellets(that she infected virus) with SDS in 100'C for 5 mins.

The thing is that she didn't mix the pellet with SDS by pipeting before she boiled it.

And to my surprise, it turned out to be a great mistake as the pellet somehow got in the shape and never is soluble at any rate; she tried everything to "destroy" the pellet like pipeting, vortexing, and flicking.

I don't know the full procedure of her experiment, so I might miss some obvious missing step, but nothing seems that wrong to deserve this unchangeable condition of pellet.

What do you think? How come this happens? Could possibly she make it soluble again?

Sorry for lack of detail and also for my broken english. :P


Thx!!


Could she still "destroy" the pettel before she boiled it?

Sometimes you can have too much material so that the pellet wont be "destroyed".

ex. when extracting dna or plasmid dna, if you do to much of the solution in 1 tube and this spin it to much to pelet it, you can get a pellet that is to big, to strong.

If you don't know it, then ask it! Better to ask and look foolish to some than not ask and stay stupid.


#3 miBunny

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Posted 01 September 2009 - 04:55 PM

I wonder if she basically "boiled an egg". The proteins aggregated together in a clump (like what happens to an egg white at high temperature)

#4 eberthella

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Posted 01 September 2009 - 05:36 PM

I'm with the bunny - it's a clot of denatured protein

#5 iamdhan

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Posted 02 September 2009 - 05:48 PM

Thanks guys!

She prepared new samples, and this time she made sure to boile them after appropriate times of pipetting. No aggregation. :P I didn't know how important this step is.




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