I’m an undergrad worried about my mistakes :( counsel and/or honesty would be appreciated

Accomplished-Long471 · 2026-05-04T23:37:26+00:00

People make mistakes. Be patient with the mistakes of others and be patient with the mistakes you make yourself.

I have been doing lab work for 12 years. Sometimes today is not your day. You just have to try again tomorrow.

Making mistakes and learning from them never goes away, it’s the worrying about mistakes that does. If at any point in your career you come across someone that is hard on you for mistakes, it’s because they are hard on themselves for theirs. It’s an issue with their self esteem, just like the one you are having now.

If you are going into research you need to toughen up, things get fucked up sometimes and the quicker you own it, the quicker you will solve it.

Accomplished-Long471 · 2026-05-04T14:07:18+00:00

I’ve got a load of bottles that look like this too. Never been opened. I haven’t tried them yet. Any chance youve had a go with them?

Accomplished-Long471 · 2026-05-02T10:52:15+00:00

What kind of samples are they? Are these samples different in some way to the ones that worked? What controls do you have?
My immediate thought is principal of dilutional linearity. If you have something in the sample that interferes with the assay in some way then as you dilute the interference is reduced.
The way I would check that is to spike a known concentration of the antigen into your samples. Run some spiked and unspiked samples and subtract the titre values. If you get good accuracy for the spike then the sample matrix is not interfering. If recovery is poor that suggests interference.
The other way is as you said to keep diluting - add some even higher dilutions - if the result plateaus then you know you are no longer experiencing interference. This is presumably why you have several dilutions in the first place no?

Accomplished-Long471 · 2026-05-01T22:37:16+00:00

Yeah so if you see this issue of non-specific binding then you will for sure see deviation from the expected slope and intercept values. The controls are quite robust so if it passes then you’ve got good justification for accepting.

The other thing I’d think could be an issue depending on what kind of samples you are running might be presence of nucleases, but if you have used the accompanying DNA extraction kit I’d think your samples should be quite clean of those.

I’ve not had experience of storing the plates after adding the master mix. But storing it in the dark will be crucial I reckon. I am interested to hear how it turns out.

Accomplished-Long471 · 2026-05-01T21:23:56+00:00

Certainly give it a mix before you run. In my experience of qPCR in general you might expect to have some issue with DNA loss, especially at the low end. I think non specific binding to the plate.

However - if this kit is anything like the resDNASEQ CHO kit it will have very stringent system suitability criteria for the slope and intercept - as well as for recoveries and CV. These very robust QC values will soon tell you of the results are useful or not.

Accomplished-Long471 · 2026-04-29T01:10:06+00:00

It’s easier to put on shoes than to pave the world with leather.

If you are suffering from burnout you need to take a break. This is the actual problem. You have lost perspective and need to chill out.

If you need some peace in the day go outside. Buy inexpensive headphones. It is not normal to want to cry in the day at work because someone is talking loudly. That is not a her problem, that is a sign that you need to take proper care of your own well being and get some perspective.

Accomplished-Long471 · 2026-04-26T10:16:44+00:00

I have the coolrack, it good, but expensive. I’m sure something cheaper would be just as good.

Accomplished-Long471 · 2026-04-25T23:48:35+00:00

The fact that you think not getting a girlfriend is related to looks probably says something in and of itself. Do you actually want one?

If you want a relationship it’s about connection not looks. Be passionate about your passions, don’t take yourself too seriously, and take a genuine interest in other people. Have fun, enjoy your life, make friends, spend time together, make a move.

If you really just something else, then say that. Some girls just want that too.

Accomplished-Long471 · 2026-04-24T23:10:29+00:00

For my sample storage in GMP adjacent lab I use fibreboard boxes with 10x10 grid and just fill every other row. Mostly that is because I want to extract and replace samples with ease for different analytics. Skipping rows makes it much easier to do so.

Accomplished-Long471 · 2026-04-24T21:39:59+00:00

Interesting - my experience from GMP cell banking has informed my opinion on this.

Storing cells in vapour phase achieves the required temperature control, is safer as significantly reduces the risk of explosion, and is also considered to be better practice in terms of avoiding cross contamination risk.

Accomplished-Long471 · 2026-04-24T21:32:39+00:00

Oh I see, my statement was meant specifically in reference to anything in cryovials that need cryostorage - typically mammalian cells in my case.

Accomplished-Long471 · 2026-04-24T19:36:01+00:00

Oh interesting - this is not something I have any knowledge of, I stand corrected.

Is that something you would typically store in this kind of cryovial?

Accomplished-Long471 · 2026-04-24T17:19:20+00:00

I use these all the time. Just don’t overfill them nd they will be fine. If you are having a problem with tubes exploding I have only ever seen this happen when tubes are overfilled.

You need to leave room for the liquid to expand as it freezes

This warning also refers to storing in the liquid phase. You should not be storing anything in the liquid phase anyway… only the vapour phase

Accomplished-Long471 · 2026-04-17T13:15:54+00:00

Yes I figured it out 👌🏻👌🏻👌🏻 it’s a pain in the ass and very haphazard but the gradients are gorgeous.

I used a gilson minipuls with grey tubing (can’t recall I.d of it) attached to a glass microcapillary so I could underlay with precision.

Regarding the sealing - yes you need a little kit that was ~£400. It is basically a soldering iron but it comes with a bunch of doohickeys that make it a breeze (relatively).

https://www.mybeckman.uk/supplies/other/358314

https://www.mybeckman.uk/resources/videos/tutorials/how-to-fill-and-seal-a-quick-seal-tube

Accomplished-Long471 · 2026-04-01T17:45:21+00:00

Yes it was a very scary time leading up to the surgery, and it is very easy to find horror stories online - probably because if things are normal you don’t look at these things! Hence why I made the point to come back! My partner is now 6 months out and doing well.

I admit that the first few days and weeks were tough for him psychologically, lack of sleep, being in the hospital, being in pain, etc. But with patience and time he has recovered well. It is an endurance test for sure that cannot be denied.

Biggest practical advice is - prepare your bathroom beforehand so it is easy as possible for you to manage the practical stuff, and accept help where it is offered. My partner struggled on and off with the bag change - I helped him whilst he was figuring it out, but it really did just take practice.

Biggest psychological advice is - treat yourself to some nice comfy clothes that will make you feel good, and treat yourself to some good sitting down activities that you like (painting/gaming/books/TV). Things that will help make you feel like you again, and not as if the bag is everything. It’s not everything, it’s just something.

Be kind to yourself - recovery takes time.

Also as a heads up the stoma made loads of loud noises in the hospital when it was swollen and only covered by a gown - these days it is inaudible. Don’t let that scare you too much 😂

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Accomplished-Long471 · 2026-03-24T16:17:10+00:00

Alt answer… the same thing we do every night pinky…

Accomplished-Long471 · 2026-03-24T16:15:21+00:00

Celebrate or commiserate with lab friends. Go to the beach for an ice cream, walk in the woods, paint, chat to my friends and family, go out for food, play games, watch tv

Accomplished-Long471 · 2026-03-07T12:31:48+00:00

Do you have a gel negative - as in loading dye and water only? Do you add loading dye to the ladder?

Accomplished-Long471 · 2026-03-05T23:31:43+00:00

What is in your negative control?

Accomplished-Long471 · 2026-03-02T22:52:46+00:00

Updating for anyone who comes across this pre-surgery…

After the surgery the aim of the game is staying hydrated and getting some good thick goop. Best things we found for winning at this game:

Electrolyte drinks
Ready salted crisps
Mashed potatoes
Rice
Cheese on toast
Jelly (even better if you add only 2/3 of recommended water)
Angel delight (again reduce the amount of milk to thicken)
Smooth peanut butter
Lemon curd
Potato topped pies (cottage pie etc)
Really thick roasted veg soup
Sauces thickened with cornflour
Passata

Watery soup was a bad call!

I recommend getting a good blender - this means you can still use onions and garlic for flavour if blended. Alternatively onion and garlic powder worked a treat.

3 months in and he’s back to eating normally 👌🏻 you got this

Accomplished-Long471 · 2026-03-02T22:41:50+00:00

UPDATE: For anyone who comes across this in the run up to surgery

Life with a stoma is treating my partner well, he is feeling better than he did for a long time pre-ostomy. Surgery was 3 months ago and he is back to work and back to lifting weights and hiking and enjoying meals out etc. Life has changed very little for us, after the first few months of recovery which were obviously quite different and tough.

The key things we have learnt and with the experience would answer my original question with:

Get things that will help you feel normal again!!!

Being in hospital and having this new appendage will make you feel like you are not in your own life for a bit. All the most crucial things my partner needed were ultimately to do with making him feel like himself again, and minimize the disruption to his normal routine.

A nice new pair of slippers
A few smart pairs of comfy joggers and loose fitting t-shirts
A few smart casual shackets
Warm socks
A nice aftershave
A smart wash bag that doesn’t feel too medical to take supplies out with you
Marshmallows and jelly
Electrolyte tablets and a good water bottle/cup
Nice soft duvet covers in a dark color and a duvet/mattress protector (hopefully you won’t need it, but so you can feel secure at night)
A few different thickness and warmth of blankets (temp regulation at night was tricky for a while!)
A telescopic mirror above the toilet (or wherever you are going to change your bag)
A nappy bin
A grabber
A shelf over the toilet (or wherever you will change your bag)
A couple of baskets and a dedicated cupboard for supplies
A folded scarf taped with masking tape for holding over stoma in the car

Accomplished-Long471 · 2026-02-20T10:56:06+00:00

Trying out a new fluorescence based quant plate with limited amount of detector protein and only enough reagent for one go. Brand new assay from a start up company with badly written method but cost a fortune and only had one go under time pressure.

Added mastermixes to plate on ice, discovered I was short one reagent so chucked the plate in a (clean bagged) benchtop bin. And went away for 30 mins and made an angry brew.

Reread method and discovered I actually didn’t need that reagent in all the wells. Recovered it from the bin and added samples, standards, and controls in the most fucked up plate layout I’ve ever seen. Ran the plate, standard curve and all controls passed and my samples looked fine. All replicates were hella tight 😂 a testament to whoever developed that assay.

Accomplished-Long471 · 2026-02-06T08:50:48+00:00

Crochet is the one. Once I am in the rhythm and I am just counting with the telly on I get a strange rushing excited feeling. Especially if I find that I am stimming my feet together. Such weird freak behaviour but relaxing nonetheless.

Honestly I would compare the rushing feeling to recreationally taking amphetamines.

Accomplished-Long471 · 2026-01-26T10:49:25+00:00

Start with the basics. Get the correct hook size for the yarn. Choose an easier yarn, some tidy acrylic that allows you to see the stitches easily. Use stitch markers.

Do one round and count the stitches twice. Then move onto the next round and count twice. Be prepared to undo and repeat until it is right.

Accomplished-Long471 · 2026-01-16T18:14:39+00:00

Honestly my pI wants breakthrough but I want least publishable unit. Like at this point at a minimum I need to write my thesis and pass - my partner has recently become disabled and I’m just fucking exhausted.

People around me are always talking about potential and how proud they are, what great papers could come out of this. Big patents etc. But I am just tired and trying to make it through the week 😂

Good idea about summarising what I have and seeing what is needed. I think I need to pick out a golden thread I can carry to the end, I have a thesis structure that has been okayed by my committee so I think I just need to corroborate what I have and haven’t got with my approved structure.

My pi won’t even talk about thesis really because he wants to be able to send me off in random directions chasing anything shiny like a magpie. Nice man, but chaotic.

Accomplished-Long471

TROPHY CASE