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Help needed on agarose gel questions

Help needed on agarose gel questions - Agarose Gel Electrophoresis Forum

Help needed on agarose gel questions - Agarose Gel Electrophoresis Forum. A forum to discuss electrophoresis staining, troubleshooting, and development of films for DNA and RNA gels.


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  #1  
Old 12-06-2011, 05:32 PM
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Default Help needed on agarose gel questions



Hi,

Can someone please tell me if I am doing the right calculation here.

So, the question is if had to make 50ml of a 2.0 % agarose gel, how much agarose would I need to weigh out?


So I find 2% of 50ml:-

2.0x50ml = 100 g of agarose ? is this correct ?


And I need to calculate how much of a 6 X loading buffer I need to add to my PCR sample, but I am not sure how to do that. Can someone please help? Many thanks
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Old 12-07-2011, 08:46 AM
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Default Re: Help needed on agarose gel questions

First question answer
To prepare 2% gel you have weigh 2g of agarose in 100ml of Buffer,

so for 50ml 1g.

Second question answer

you have to add 1ul of 6x loading for 5ul PCR sample
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Old 12-07-2011, 11:40 AM
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Default Re: Help needed on agarose gel questions

The way I remember how much sample buffer to add is like this:

Whatever the concentration of the sample buffer is (say 10x) you divide the volume of your sample by 1 less than the concentration (9).

For instance if you have 23ul of sample you need to add:
23 / 9 = 2.55ul of 10x buffer

You can check it:
23ul Sample + 2.55ul 10x sample buffer = 25.55 ul total volume (this would make the buffer 1x which is what you want)

So for your example, divide the volume of sample by 5 to get the amount of 6x buffer you need,
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Old 12-13-2011, 06:16 PM
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Default Re: Help needed on agarose gel questions

So if i had to make a 1% agarose gel of 50ml how much agarose would I need and how much of TBE ?

and if it was 1% but double the amount (100ml gel) then how much of agarose and TBE would i be adding together?
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Old 12-14-2011, 12:35 PM
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Default Re: Help needed on agarose gel questions

Hi Gamewizard,

I already replied that if want to make 2% gel you have weigh 2g of agarose in 100ml of Buffer.

Just downgrade the for 1%:

1g in 100ml or 500mg in 50ml.

And i didn't understand your second question.
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