0
$\begingroup$

I want to load my agarose gel with my PCR products, which are 50 µL reactions.

I am wondering how much of those reaction I can load into one well if I use a standard agarose mini gel (10 well comb, ~8 mm thickness, 1.2%).

Should I try loading the entire 50 µL into one well or should I split it into two?

Thanks in advance

Phil

$\endgroup$
  • $\begingroup$ Why do you want to load so much sample? $\endgroup$ – Chris Jan 13 '17 at 10:39
  • $\begingroup$ Low concentration or for gel extraction I would assume... $\endgroup$ – Joe Healey Jan 13 '17 at 11:03
  • $\begingroup$ Yeah, it is for gel extraction. I want to load maximum sample while using the minimal amount of gels $\endgroup$ – Philz0rs Jan 13 '17 at 11:30
  • $\begingroup$ If this is for gel extraction, load two lanes. Or connect two on the comb with tape to make a bigger well. $\endgroup$ – Chris Jan 13 '17 at 11:47
1
$\begingroup$

How many samples have you got? You can tape neighbouring teeth of the comb together to create double- or triple-wide wells to load more sample.

I don't normally load more that about 20 µL.

You could also make the gel thicker potentially.

$\endgroup$
  • $\begingroup$ Thanks for the tip! Unfortunately, I only have one standard agarose apparatus, and it doesn't allow me to use wider gels. I tested with loading buffer in one well, whether or not it would fit 50 µL. It did, but I didn't take the risk of losing sample due to leakage, so I will just run more gels. Thanks a lot, though! $\endgroup$ – Philz0rs Jan 13 '17 at 11:32
  • $\begingroup$ @Philz0rs If this answer addressed your problem, please consider accepting it by clicking on the check mark/tick to the left of the answer, turning it green. This marks the question as resolved to your satisfaction, and awards reputation both to you and the person who answered. If you have >= 15 reputation points, you may also upvote the answer if you wish. There is no obligation to do either. $\endgroup$ – MattDMo Jan 13 '17 at 12:58

Your Answer

By clicking “Post Your Answer”, you agree to our terms of service, privacy policy and cookie policy

Not the answer you're looking for? Browse other questions tagged or ask your own question.