I picked up a Cal 2-29 recently, and I'm trying to figure out what all the parts of the head are and how to operate it. The tubing on the far left with the hand pump seems to pump from the holding tank to the waste port on the deck, and the holding tank is in the cabinet behind the head. I tried a couple different combos to get the bowl to fill with water or flush, but I was unsuccessful, and I don't know if I'm doing it wrong or it needs to be repaired.
Nonsterile may not be too much of an issue, but if you want to grow the cells in these plates, it may be worth it to make sure they're TC treated. You could also grow the cells elsewhere then lift them and do the read in the new white plates, but this can also introduce variability.
If you're looking to work with in fusion now, here's the protocol I tend to use for the whole process. Some of it is specific to where I get primers and sequencing, but the principles would apply across the board. https://drive.google.com/file/d/1FxJVhH_apvrY_7BjDZRBSpneDGfwS1Ju/view?usp=drivesdk
Which program? In Tetrad we were given new Macs in my year.
What screen protector worked for you? The first one I tried wasn't a good fit for this watch.
You could try a pipe/tubing cutter. I've used them for PVC tubes nicely before. It's likely overkill for this application, but it wouldn't require heating the plastic.
I've tried NaN3 before, but the results were disappointing at first as OP describes. I wasn't up for optimizing it when 30% H2O2 worked really well for me right away.
You can just hold it. I've commuted three years on the Richmond Ferry and have always found a spot. I'd definitely recommend a bike cover as every bike storage location will let salt spray onto the bike. I've seen actual waves come over the back of the Bay Breeze ferry and soak carbon bikes.
And it was actually named after a person, Edwin Southern! The rest of the cardinal blots followed as a pun.
I really like Topeak's Ratchet Rocket Lite for a similar tool.
I usually run a lane or two of Laemelli/Sample Buffer on both sides of my gel if I have space. That adds the same buffer to the outer wells and let's it spread out instead of the samples.
It's not immediate, but you are correct in thinking that you don't need to do anything else after transfecting to get expression if your plasmid contains a constitutive promoter like CMV.
You can bike into Live Oak and Juniper Campgrounds on Diablo. Not sure if that's what you're looking for though.
It's extra work, but you could calculate the leftmost values of the range you want to show and then graph (but hide the legend entry) an extra set of white/background colored on the left to move your stack further out.
You can always redo it and try transforming both to give yourself a reference in the future as to whether or not it's likely to work.
In the guide they mention that overincubation leads to aggregation of the DNA. If you have the materials on hand to redo it, it's likely worth it to make sure it has a better chance of transforming well.
It's not the same kit, but Qiagen's P1 resuspension buffer is: 50 mM Tris-HCl, 10 mM EDTA, pH 8.0 (25ºC), 50-100 µg/ml RNase A
One way I've done this is with a split luciferase assay (NanoBiT). We transiently transfected two plasmids into HEK293T cells: one expressing CAAX_LgBiT, the other SmBiT_POI.
More likely, if the biker was further over, they would have had the same problem, and even more vehicles would be passing too close. I have somewhere around 15-20,000 miles of bike commuting experience, and riding further to the right simply encourages more closes passes. It's a night an day difference riding on the very right vs. taking the lane as allowed. Additionally, when you're already all the way over to the right and someone close passes you, there isn't anywhere to go when trying to escape them. By riding to the left, you leave yourself more space to escape a close pass into.
Here's one from Promega where they compare with forward transfection. https://www.promega.com/resources/pubhub/reverse-transfection-using-fugene-6-and-fugene-hd/#:~:text=In%20reverse%20transfection%20protocols%2C%20cells,compared%20to%20standard%20transfection%20protocols
Are you referring to the grey value at the y junction that has been zip tied?
Could someone help me identify what the different tubes in this marine head are and/or how to operate the different functions?
sailing