3 replies to this topic

[PhD]

Its me again, gees it seems to be Bioforum day with me.

Im looking into trying electroelution to get rid of one high MW contaminate I have in my eluted fractions but cannot affort an electroeluter.

How can I set it up just used my dialysis membrane? I run my gel in mini protean tetra cell from Biorad?

Neone have a detailed protocol for this kind of thing? I would really appreciate it!!

[swanny]

PhD, on Aug 24 2009, 10:01 PM, said:

Its me again, gees it seems to be Bioforum day with me.

Im looking into trying electroelution to get rid of one high MW contaminate I have in my eluted fractions but cannot affort an electroeluter.

How can I set it up just used my dialysis membrane? I run my gel in mini protean tetra cell from Biorad?

Neone have a detailed protocol for this kind of thing? I would really appreciate it!!

What's the rest of your protocol, please?

[tfitzwater]

Electroelutions were originally performed in an agarose gel box.  Clamp the bottom of a piece of dialysis tubing and fill to the brim with buffer.  Drop in the gel slice and squeeze out most of the buffer.  Avoid air bubbles that result from eliminating too much buffer.  Clamp the open end of the dialysis tubing and lay the slice down on the gel box.  Add buffer to cover slightly.  Measure the distance between the electrodews and run at 2 volts/cm.  If the gel slice was prestained, you can monitor the elution of the DNA/RNA out of the slice.  Before harvesting, reverse the polarity for 30 seconds to back the DNA/RNA off the membrane.

[PhD]

Ola!

thanks for the replies I actually had no protocol since I asked around in lab if neone new how to do it and I was discouraged from doing it. It doesnt seem complicated at all! The only thing that i wonder is how much electrophoresis buffer to put in the bag so protein is not too diluted? what would be the minimum amount for a slice of gel in approx 8cm bag?


I will give it a shot!


have a great day!!