- #1
MatthewHaas
- 15
- 0
Hi everyone,
I am running agarose gels (1.5%, 100 V, 100 min) with the product of PCR-RFLP.
My gel has 96 wells...48 on top and 48 on the bottom, so it is a rather large gel...300 ml by volume.
I am having trouble getting clear, bright bands for the DNA in the lanes near the edges. It is very faint. When I shift the gel in the UV box, so the edge is closer to the center, I can see the bands very clearly.
Do I need more EtBr to see this when the gel is in the position I want to photograph? I added 15 ul initially, but reuse the gels (per instruction) and out of fear for overloading the gel, I don't add EtBr each time even though I think it is being degraded when I microwave the gel to get it into a liquid again. When I do, it is on the order of 2-3 ul.
I am running agarose gels (1.5%, 100 V, 100 min) with the product of PCR-RFLP.
My gel has 96 wells...48 on top and 48 on the bottom, so it is a rather large gel...300 ml by volume.
I am having trouble getting clear, bright bands for the DNA in the lanes near the edges. It is very faint. When I shift the gel in the UV box, so the edge is closer to the center, I can see the bands very clearly.
Do I need more EtBr to see this when the gel is in the position I want to photograph? I added 15 ul initially, but reuse the gels (per instruction) and out of fear for overloading the gel, I don't add EtBr each time even though I think it is being degraded when I microwave the gel to get it into a liquid again. When I do, it is on the order of 2-3 ul.