Ignore the EvoSep instructions and load peptide tips however you want!!

The whole point of the EvoSep HPLC is reproducibility, I'm pretty sure. That's what it says on all their things. The tips come with handy little visual instruction cards that tell you exactly how to load them. My team actually made stickers for the boxes where you check off each step you've completed because we can only spin 2 boxes at a time in our centrifuge and we often prep far more tips at a time. 

And...I swear about half of the papers I read (particularly from one group I'm going to point out right now) seem to just load the tips however they happen to feel like doing it that day. 

This is by far the funniest one. In this paper on AlphaDIA - a group that has strong ties to EvoSep - ignores the instructions entirely - in 2 separate ways, in a single paper. 

Look, this is funny because it's the same study, but if the goal is reproducible peptide binding why introduce a potentially confounding variable at all?? 

First EvoTip prep

Propanol-1 then water then 99% acetonitrile then re-equilibrate (you never learn how much volume here).

And then a totally different way of loading the same tips! This time with a robot, so maybe it has to do it this way? 

Propanol-1 then 99% acetonitrile twice then water twice with seemingly random volumes? 

In case you aren't familiar this is what the card says in the box. 

It was tough choosing a gif here, but it's tough to pass this one up.