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2X sample buffer vs. 6X


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5 replies to this topic

#1 Princesslinda

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Posted 20 July 2009 - 04:40 PM

What is the difference between 2X vs. 6X sample buffer and how do you know how much you should add of each into your samples before loading your gel? Thanks!

#2 jiajia1987

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Posted 20 July 2009 - 05:25 PM

THe difference just lies in the concentration.

2X means you need to do a 2X dilution. So say.. you wanna load 10ul, that will be 10ul/2 = 5ul of your sample buffer to load to 5ul of your samples.

6X means a 6X dilution is required, so say... for 10ul, it will be 10ul/6 = 1.7ul of your sample buffer to load to 8.3ul of your samples.

#3 Princesslinda

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Posted 20 July 2009 - 06:55 PM

so for example, if I have a stock that has a concentration of 10 micrograms/microliter and I want a concentration per lane of 100 microgram/microliter...I would need 10 microliters to get this concentration. How much 6X would I need to add to this?

#4 HomeBrew

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Posted 20 July 2009 - 07:40 PM

Add 2 ul to 10 ul. It's close enough -- you need not worry that your loading dye is not precisely 1x, just that it is sufficient to serve its purpose -- to weigh down the DNA so it won't float out, and give you an observable marker for the run.

#5 mikew

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Posted 21 July 2009 - 12:20 PM

The difference is 4x :D.

#6 leelee

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Posted 21 July 2009 - 11:07 PM

It means how concentrated it is, ie how many times (hence the X) working concentration (or 1X) it is.
So, for example, your 2X buffer is 2 times more concentrated than a working concentration of the buffer.


PS.

The difference is 4x smile :D

hahaha good one mikew!!




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