E-Gel Size Select

[sflanagan]

Hi there,

I'm looking into purchasing an E-Gel Size Select from Life sciences, and was wondering if anyone already has one. I would like to know if you have any comments, criticisms, or rave reviews about it before I buy one. Anything is useful!

Thank you!

[hyjkim]

I've used one. It's great for selecting very specific pcr bands, but not good for selecting ranges of DNA. If you're trying select a range for NGS then, I'd suggest you check out the Sage Science PippinPrep or the Caliper LabChipXT.

[Simone78]

I have been using it HEAVILY in library preps for methylome seq (2% gels). Just one comment: it´s great! The range selected is narrow, which is good if you are interested in selecting fragments with approximately the same lenght. It´s very reproducible, fragments of the same size migrate into the collecting wells after the same time when run on different gels (+/- 30 sec, seldom 1 min). The run can be monitored in real time, so this is not really an issue.
As a backup, during library prep I was collecting several fractions at 30 sec intervals and it was very easy. Just stop the gel, recover your DNA from the collecting wells, refill it with water, run the gel another 30 sec, stop it again and collect another fraction. I was then performing bis-conversion and PCR on a single fraction. If something went wrong I had other aliquots as a backup.
Attached is an example of a typical profile after size selection with 2% E-gel.
good luck!

[pbluescript]

I agree with Simone78. I use either Ampure beads for size selection when I don't need a tight band or the E-gels when I need a more specific size. I do a very similar protocol and select multiple size ranges. I have noticed that different gel lots will run at different speeds and for some reason the library sizes I collect are smaller than what my ladder would predict. However, if you collect a range of sizes, you'll get what you want. It's a nice alternative for those who don't have the budget for a PippinPrep or Caliper.

[abcdef]

Hi there,

What is the typical recovery rate using the E-Gel Size Select?
Has anyone tried to work that out?

Thanks!