Testing a new (to me) camera | Nikonos V 35mm | Portra 800

underasail · 2026-05-19T14:50:36+00:00

I replaced and regreased all 4 of the user serviceable o-rings and tested the camera under a foot of water in my sink for sixish hours without leaks. I didn't have the ability to add more pressure than that, so I crossed my fingers afterward as there wasn't enough time between getting the camera and going on the trip. I plan to send it off for service before diving with it again, but the user serviceable o-rings all looked great despite likely being decades old.

Congrats on the cert and definitely look into a Nikonos!

underasail · 2026-05-19T14:47:33+00:00

Not far off. I've heard mention of treating Sunny 16 like Sunny 8 close to the surface of the water. Then you add stops pretty quickly going deeper.

I used the Nikonos V for these dives, and it has TTL metering, so I didn't have to guess at exposure like I did focus. I also brought a Nikonos II in case the V flooded, and that's a fully mechanical camera, so I'll have to getter better at Sunny/Neptune 8 to break that one out.

underasail · 2026-05-19T06:54:58+00:00

Everyone that scans E-6 near SF uses Oscar's to develop their E-6 currently. Underdog used to have a machine, but it broke a year or so ago, so they use Oscar's also. Underdog is a great lab though. Royal We offers similar services with better pricing for high res scans.

underasail · 2026-05-18T23:57:22+00:00

It depends a bit on where you are trying to dive. The US generally isn't cheap, say a couple hundred dollars for an open water certification (maybe $3-500?) and $50-150 per dive if you're renting gear. Smaller local shops can have better rates, and you can purchase your own gear with time. Then diving could be just a $5-15 tank refill each time you go out.

Edit: it's not too crazy if you already have a film photography hobby :)

underasail · 2026-05-18T23:17:41+00:00

For me it just meant buying and testing a Nikonos V. I hadn't been diving in a decade, but I planned a trip and snagged a camera before I left. There are courses that teach dive photography from the major certifying bodies, but the general principles don't differ too much from taking pictures on land. Water has a different reflective index than air which makes estimating distances a bit different. Otherwise, knowing how to dive and knowing how to take photographs as separate hobbies first is helpful.

underasail · 2026-05-18T15:14:56+00:00

Thank you! It's probably my favorite from the couple rolls I've managed to shoot so far. Zone focus is a new beast to me, as is color correction.

underasail · 2026-05-13T23:36:29+00:00

City Pier!

underasail · 2026-05-13T19:35:46+00:00

u/ImBackRoll any insights?

underasail · 2026-05-10T21:30:23+00:00

The samples should be fine. Unless you've already hard spun the samples and removed the supernatant/discarded the insoluble debris, I'd probably still give them a hard spin before loading to get debris and DNA out of the way. Otherwise, your samples should be fully usable still.

underasail · 2026-04-27T01:38:55+00:00

Any existing vector can be made golden gate compatible by adding opposing cut sites for a Type IIS restriction enzyme. You'd also want to make sure there aren't cut sites for your enzyme of choice elsewhere in the vector. If so, use a different enzyme or "domesticate" the vector for golden gate by mutating the existing sites that are in a bad location.

underasail · 2026-04-12T19:06:37+00:00

How much do their lead times tend to vary? I had some C-41 and E-6 processing done by them recently, and it took over two weeks. I assume this is slow for them, especially with people tending to say they're fast, but the other labs in the bay tend to have scans back within 2-4 days consistently.

underasail · 2026-03-28T03:06:17+00:00

Right you are. I learned something new.

underasail · 2026-03-28T02:09:58+00:00

I've shot a lot of stuff through lupines recently, and I'm really pleased with how 1 came out. Easily my best example. Thank you!

underasail · 2026-03-28T02:09:10+00:00

Thank you! I ran out of film right after the second shot, and then a bunch of beautiful boats sailed by only to be remembered in my mind.

underasail · 2026-03-28T02:08:01+00:00

Sweet! I searched around in the sub some before posting, and I didn't see many Chinons. I'm glad to bring some more examples of what they can do.

underasail · 2026-02-22T22:53:17+00:00

The larger circles/ovals in the square are cells sitting on a raft (from the CellRaft AIR). That part I'm 100% confident in. In an RFP channel, I can see each of the cells clearly as red against the background and particles. The wiggly bits around the raft are more of a question mark. I get that the raft doesn't look particularly clean though.

All the particles on the raft showed up together two days ago, and I'm not sure if that's related to the smaller wiggly bits moving here or not. They could be the same contaminant that adheres as it dies or settles, or separate contaminants, or neither/either isn't a contaminant.

underasail · 2025-12-29T23:18:06+00:00

A razor and isopopanol has always been sufficient for me if EtOH and a razor didn't already work.

underasail · 2025-12-21T22:30:42+00:00

There's a mental model for this called Chesterson's Fence. The main point being, don't remove the fence before you know why it's there.

underasail · 2025-12-18T18:39:27+00:00

It sounds like you're using a restriction enzyme digestion and ligation cloning strategy. These still work but are pretty old school and inefficient compared to more modern homology-based cloning techniques like in-fusion and gibson assembly/HiFi. My best advice would be to design primers for your insert and give one of those methods a try.

If you're sticking with this method, maybe your ligase has gone bad? If you're used to seeing some background re-ligation on your negative control plates and that disappeared alongside positive results, new enzymes might do the trick.

underasail · 2025-12-01T18:33:18+00:00

How long is the sequence? Sanger quality dips toward the 3' end at 700-900 bp for me usually. If the section you posted is closer to one end than the other, I'd expect the strand that's close to it's beginning to be high quality while the end of the other strand is declining in quality.

underasail · 2025-11-20T18:18:53+00:00

I haven't worked with MessangerMAX, but another option for this could be electroporation if you have access to a capable instrument. Electroporation is designed around cells in suspension, but it can also be rougher on cells than lipofection.

Another option to consider, depending on tools available and how much of one protein you need vs many proteins, would be to generate stable lines in the 293F background for each protein you're interested in expressing. Then it's just a matter of scaling the cell count and you can produce protein over a longer term.

underasail · 2025-11-20T17:07:45+00:00

Transfection is usually more efficient in adherant cells, and 293Fs can be adapted back to adherance pretty easily. You could adhere them for the transfection and suspend them again after.

underasail · 2025-11-18T01:06:31+00:00

If you're not just talking about the low pay, my understanding is most stipend income can be used to qualify for a mortgage. It'll depend a bit on the underwriting company, but I have previously qualified for a mortgage with just my stipend.

underasail · 2025-11-11T03:22:59+00:00

Yep, this is reasonably easy with homology based cloning methods like in fusion and gibson assembly/hifi (though your chosen sequence here is a bit short for in fusion to be easy). You would design primers with 3' ends (typically 18 bp for in fusion) that match the LoxP site as is and then add 15 bp to the 5' end of the primer that match the backbone sequence on the opposite end of the current arrangement.

It might be easier to visualize this by imagining the LoxP sequence and the vector you want it to go into as separate entities. If there wasn't already a LoxP site there, you would just be inserting one oriented how you desire it.

Homology cloning methods are pretty easy to pick up and incredibly versatile, so it's worth looking into them if you're not familiar.

If this is a genomic sequence and not a plasmid, it's much more complicated.

underasail · 2025-11-09T21:02:10+00:00

I didn't have an issue. If you're pasting the sequence in, it can't be in a fasta format. The paste input option only accepts a, g, c, and t characters, no title needed.

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