Anonymous

Electrophoresis Stains: Difference between revisions

From London Hackspace Wiki
Fixed SYBR Green in Taq discussion (it's a rt-pcr stain)
(Fixed SafeWhite error (it's per reaction, not per gel))
(Fixed SYBR Green in Taq discussion (it's a rt-pcr stain))
Line 59: Line 59:
Three comments on all this:
Three comments on all this:


# Even though it's really cheap, I don't like SYBR Green in readymix. The problem is:
# Even though it's really cheap, I don't like SYBR Green in readymix. It's designed for RT-PCR, which we're not doing at the moment. If we don't use it for RT-PCR, then we will end up visualising a ladder using a different method from the one we use to visualise the samples, which sucks: we won't know if we screwed up the visualisation or if the reaction didn't work (this problem is what led us to ethidium bromide in the first place).
## I'm all about reducing the number of variables per reaction.
## We can't use SYBR Green + Taq to visualise ladders directly
## Because we won't be running a ladder mixed with the same dye (at best, we'd mix the ladder with SYBR green after PCR), we won't know if we screwed up the visualisation or if the reaction didn't work (this problem is what led us to ethidium bromide in the first place).
# Even with disposal costs, ethidium bromide isn't that bad. It's less expensive than SafeWhite (for 2 or more reactions per gel), but, to be fair, that's still not a significant difference (10p or so). If we handle EtBr properly and dispose of it correctly, we should be fine.
# Even with disposal costs, ethidium bromide isn't that bad. It's less expensive than SafeWhite (for 2 or more reactions per gel), but, to be fair, that's still not a significant difference (10p or so). If we handle EtBr properly and dispose of it correctly, we should be fine.
# For £40, we may as well check out SafeWhite. It's not particularly cheap, and we will still have to retain LabWaste for waste disposal, but we won't need them to dispose of gels, so we won't need them to come as frequently.
# For £40, we may as well check out SafeWhite. It's not particularly cheap, and we will still have to retain LabWaste for waste disposal, but we won't need them to dispose of gels, so we won't need them to come as frequently.