PAGE with Realtime PCR product (Sybr Green) - Post-staining with ethidium bromide? (May/12/2008 )
hello! in addition to melting curve-analysis i want to check the product of my realtime pcr (sybr green) on an polyacrylamide gel.
now i'm wondering if the residual sybr green in the dna does interfere with EthBr post-staining of the gel (as for instance blurring of the bands). anybody already done something like this? any suggestions are very welcome!
I can't be sure on a PAGE analysis, but on an agarose gel, it does not interfere.
My guess is that it will be okay
thank you. do you have to adapt the standard-staining procedure (EthBr) when you are using dna with sybr green? or is the flourescence of sybr green in the gel negligible?
after melting curve i make an additional step at 72°C for 10 min, then i run my samples on agarose gel and i don't adapt the EtBr procedure. It works good