I got 60 micro-litre samples from IP and want to load them into single wells respectively. The problem is that the well can only hold about 30 micro-litre. How can I load 60 micro-litre sample to a single well in SDS-PAGE gel?
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How to load 60 micro-litre sample to a single well in SDS-PAGE?
Started by felixwang, Mar 29 2005 07:45 AM
6 replies to this topic
#2
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Posted 29 March 2005 - 09:03 AM
sir,
i'm not sure about what i'm saying but i think it may be possible to load 60
micro-litre sample by increasing the the thickness of the gel.sir,once again i'm clarifying that i'm not sure about it.
srini.
i'm not sure about what i'm saying but i think it may be possible to load 60
micro-litre sample by increasing the the thickness of the gel.sir,once again i'm clarifying that i'm not sure about it.
srini.
#3
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Posted 29 March 2005 - 10:08 AM
Hi,
you can tried to load your sample on a 'big gel' (about 7'*7'). The comb of these gel allowed to load easily 60uL.
Also, you can tried to lyophilied or dialysed/lyophilied your sample. With these manip, you can probably reduced the volume of your sample to 30 ul.
good luck
you can tried to load your sample on a 'big gel' (about 7'*7'). The comb of these gel allowed to load easily 60uL.
Also, you can tried to lyophilied or dialysed/lyophilied your sample. With these manip, you can probably reduced the volume of your sample to 30 ul.
good luck
#4
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Posted 29 March 2005 - 03:24 PM
What I've done in the past (and it seems to work 
) is to load 30 uL and then run the sample towards the botton of the stacking gel and load another 30 uL before it reaches the seperating gel. You have to be 'on-the-ball'; but it's always worked for me.
) is to load 30 uL and then run the sample towards the botton of the stacking gel and load another 30 uL before it reaches the seperating gel. You have to be 'on-the-ball'; but it's always worked for me.
#5
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Posted 30 March 2005 - 12:20 AM
hi
if you use a small pa gel you can make a "double well" using adhesive between two teeth of the comb.
if you use a small pa gel you can make a "double well" using adhesive between two teeth of the comb.
#6
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Posted 30 March 2005 - 06:13 PM
I would add your denaturing buffer to your 60ul of sample then put it at 99 degrees with the lid open - this will allow the water to evaporate and thus concentrate your sample.
#7
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Posted 30 March 2005 - 11:39 PM
Nay, on Mar 31 2005, 03:13 AM, said:
I would add your denaturing buffer to your 60ul of sample then put it at 99 degrees with the lid open - this will allow the water to evaporate and thus concentrate your sample.
hem...
as your sample is soluble in water, don't you loose part of your sample too?
