in a 1% agarose gel, at 20V, the band are still very close and I'm still unable to cut one of them from
You could try to increase the size of the well, use percentage of agarose lower than 1% and use a longer gel to permit better resolution.
You could try to increase the size of the well, use percentage of agarose lower than 1% and use a longer gel to permit better resolution.
So you think that lowering the agarose concentration is a good thing?
Other people with whom I've discussed this suggested an increase, to 2 or even 3%...
I know small concentrations allow for better resolution of small fragments, but do they resolve better small differences between big fragments?
Hi Joao
like Ana...In my opinion also you should decrease the concentration of agarose gel and increase the size of the gel.....you are running gel at very low voltage...(20V for 24h)...running at very low voltage will have another problems....the separation of the fragment will be difficult due to the diffusion of the fragments also....so you need to take care of this also....mean while if you are using regular agarose ....se if other types of agarose (wide range, high resolution etc) offer some help in the range of separation you need
bye
@ Joao C
1. In my opinion you need to use higher concentration of agarose (2- 3%) for better resulation between fragments.
2. For this you can also use Metaphore Agarose which will give more resulation between bps than the normal agarose...
good luck...
Thank you all for your replies and suggestions.
I used a 1% agarose gel, around 25cm long, and ran it for 38h20m at 40V, and I managed to separate the two bands and purify the upper one.
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