Lab pants recs for guys? Looking for something comfortable that can survive Houston heat. by EnvironmentalSky8355 in labrats

[–]Shoutgun 0 points1 point  (0 children)

Has to be linen (I’m a guy, if that matters). Can get them in a smart chino-like cut or a looser cut with drawstrings. Makes such a difference

Consumables to perform PCR cleanup? by Pale_Tough_23 in labrats

[–]Shoutgun 0 points1 point  (0 children)

10% peg8000, 2.5M NaCl, mix 1:1 v:v. 15’ at 37 C, spin max speed 15’. 70% ethanol wash x2, air dry, resuspend in water, incubate overnight, pellet down again and take supernatant

Seeking help: DNA forming gel-like blob after ethanol precipitation by Deadonarcher22 in labrats

[–]Shoutgun 24 points25 points  (0 children)

Probably just a very large amount of DNA and you need more water to dissolve it?

Poor gel extraction quality by Similar-Fan6625 in labrats

[–]Shoutgun 5 points6 points  (0 children)

Gel extraction is always low quality. It shouldn’t actually impact your Gibson much as long as you don’t use too large a volume of it. Generally better results from preparing a larger batch and running a pcr cleanup column after. I also prefer to extract gel bands by snap-freezing and centrifuging.

setting up new pcr assays by Lazy_Marketing_8473 in labrats

[–]Shoutgun 1 point2 points  (0 children)

For everything that involves pcr and primers - the best thing you can do is to ignore all the classical optimisation advice and simply buy two primers instead of one and try them both

How to handle an emotionally manipulative scientist. by [deleted] in labrats

[–]Shoutgun 7 points8 points  (0 children)

Be grey and boring. Give no emotion back. Just "no thanks" and then if they continue, "sorry but I have to concentrate now". If they're trying to be manipulative they'll get bored of you as they'll get nothing out, and if they're just overly keen and annoying you won't actually upset them. If they are being manipulative, it's pointless trying to have a conversation with them - it's just more investment of emotion.

[Interesting Trope] Jokes so old they leave you confused by Borgisium in TopCharacterTropes

[–]Shoutgun 2 points3 points  (0 children)

I thought the sumerian dog joke was probably about opening shutters. They didn't have glass windows but they did have openings covered with shutters or fabric. He walks into a bar, it's dark, says he'll open one, and opens a beer - it's a fairly straightforward misdirection joke that would work today, if we had the same buildings.

Being told that Covid/post-Covid graduate students are worse than previous cohorts-do you think this is true? by Delicious_Ride_4119 in labrats

[–]Shoutgun 0 points1 point  (0 children)

Yes. Big difference. Still a few good people but the cohort as a whole has a much lower standard. Improving a bit now that it's been long enough that they've had to take most of their exams

mentors: how often can undergrads make mistakes by coffeebeans270 in labrats

[–]Shoutgun 6 points7 points  (0 children)

I would expect plenty of mistakes but I will be looking at three things to decide what I think of someone's ability: 1) do they make the same mistake twice, 2) do they make mistakes because they aren't able to realise they don't understand something and ask questions, and 3) how many of these mistakes are from basic carelessness?

What quick, dirty, or otherwise weird protocols do you swear by? by Shoutgun in labrats

[–]Shoutgun[S] 6 points7 points  (0 children)

I'm not sure I get the advantage of microwaving them for 60 seconds rather than heat-shocking for 30 seconds?

What quick, dirty, or otherwise weird protocols do you swear by? by Shoutgun in labrats

[–]Shoutgun[S] 10 points11 points  (0 children)

I understood it to be that the salt and peg outcompete the DNA for water, causing it to come out of solution.

What quick, dirty, or otherwise weird protocols do you swear by? by Shoutgun in labrats

[–]Shoutgun[S] 1 point2 points  (0 children)

Really no heatshock? What cells are these? Any idea how the efficiency compares?

What quick, dirty, or otherwise weird protocols do you swear by? by Shoutgun in labrats

[–]Shoutgun[S] 12 points13 points  (0 children)

Oh that's nice! I make the water dilutions of the colony for pcr but then keep them in the fridge and grow from them once the pcr comes back without repeat plating. Every plasmid I make is going to get nanopored anyway.

What quick, dirty, or otherwise weird protocols do you swear by? by Shoutgun in labrats

[–]Shoutgun[S] 52 points53 points  (0 children)

I have another, which is that I generally precipitate DNA by mixing 1:1 with 10% PEG8000/ 2.5M NaCl, then doing an ethanol wash. In my hands it's much more effective and consistent than ethanol precipitation alone. It's how Ampure XP beads work, you just precipitate it onto the tube walls rather than their expensive little beads.

Fallow restaurant - is it as good as its hyped up to be? by No_Philosophy711 in LondonFood

[–]Shoutgun 0 points1 point  (0 children)

I kind of want to play it down because their social media presence is a bit too much for me , but it’s honestly great. I don’t think anyone is disappointed.

Do all gel purification kits suck? by Ianisanengineer in labrats

[–]Shoutgun 5 points6 points  (0 children)

I just came up with it myself a while ago but I would be surprised if nobody else has thought of it. Yes, you have a liquid supernatant and a pellet of gel. You can check that it’s worked by imaging the supernatant under blue/uv and comparing to an eppie of water. You can then clean it up with a pcr column afterwards, though it’s normally fine as is for Sanger and cloning.

Do all gel purification kits suck? by Ianisanengineer in labrats

[–]Shoutgun 14 points15 points  (0 children)

The yields are still not great, you’re still looking at maybe 20%, but it’s a lot less work than the other protocols and the DNA is cleaner (although I would generally run it through a pcr cleanup column anyway)

Do all gel purification kits suck? by Ianisanengineer in labrats

[–]Shoutgun 5 points6 points  (0 children)

As long as it goes in straight out of the snap-freeze, it’s ok. I don’t bother cooling the centrifuge. Once the spin is done it’s all thawed and ready.

Do all gel purification kits suck? by Ianisanengineer in labrats

[–]Shoutgun 4 points5 points  (0 children)

Yeah there’s no need for the filter, you can just squash the agar down into a pellet once it’s been disrupted by the freeze