Published: September 16th, 2016 Last Modified: June 16th, 2017
Just letting you all know that I’m not dead, I haven’t been hit by a bus and I haven’t given up on science to enter the shady world of underground street fighting. I’ve been pushing hard to construct an infectious clone of a plant virus, which has been my baby for the last year and a half. I’m at the stage where I have to get the plasmid into my plants…agroinfiltration? It’s an option, but not very exciting, so instead I’ve been building various DIY particle bombardment devices!
This includes attempts at using a coilgun to shoot home made magnetite beads, an electromagnet to pull the beads INTO the leaf, and a more promising DIY gene gun that uses CO2 cartridges, and actually works!
On top of that I have articles lined up concerning DIY electrophoresis equipment, cheap digital thermometers and various useful gadgets that make our lives a bit easier. Stay tuned!
Hey! was wondering if you know the composition of the Qiagen RLT buffer used by RNeasy protocol. It seems hard to find online, but maybe you know a source? Thank you!
I don’t personally use the RNeasy kits, but I seem to have found something that could be useful for you, take a look at this patent:
https://patents.google.com/patent/US20140356860
Just taking a look at the RNeasy protocol, it seems you have to add an equal volume of 70% ethanol, which brings it in the 35% range, which is close to what they talk about ” The resulting binding mixture comprised the chaotropic salts GTC and GuHCL in an overall concentration of 2M, 33% ethanol and Triton X-100.”
This is a bit of a guess, but here’s what I would try first:
Putative DIY Buffer RLT:
4M Guanidine Thiocyante
10% Triton X-100