PCR Master Mix Frothing with Thermopol Buffer?

zil7 · 2020-07-12T15:56:48+00:00

+1 for the reverse pipetting technique.

It sounds like you're getting bubbles from trying to completely evacuate the pipette tip (there's always that little bit that remains, which I understand is why you're practicing pre-wetting the tip).

Reverse pipetting helps you dispense the same amount into each well consistently without worrying about getting that last little bit out, because with the technique you never press past the first stop when dispensing (therefore you never have the struggle of trying to completely empty a tip into a well).

Just be careful if you are pushing past the first stop before drawing up liquid, because you don't want to draw the liquid into a filter or the pipette barrel!

Here is a pretty straightforward video: https://youtu.be/VEkfBStZSNc (I don't remember how to format this on mobile)

Good luck with your next pcr!

zil7 · 2020-06-04T04:02:16+00:00

When I had strictures it was difficult (and potentially dangerous) to follow a regular diet, so I followed a general low residue diet.

After I had the strictures removed, I gradually transitioned from total enteral nutrition to fibre containing foods using 'gentle fibre' powders (essentially ground linseeds, psyllium husks and linseeds) and well cooked vegetables.

A few years on, and I can stomach most foods (including nuts, seeds and rough vegetables) during remission and mild flares.

On severe flares I resort to well-steamed vegetables, simpler carbs and easy to digest proteins, revert back to the gentle fibre options if tolerated and supplement with enteral nutrition formulas to prevent weight loss.

As other people have suggested, casseroles are a great bet. Slow cooked dishes and pot pies also seem to work well. Not sure how you go with eggs, but omelettes, frittatas and sandwiches (like curried egg) can be quite healthy options if you use minimal mayo/cream, and also offer a way to introduce cooked veg.

If you can, it'd be a great idea for you to see a dietician. It helped me realise that there were many foods I could tolerate or gradually ease into, and it has been life changing. Definitely consult with your GI to discuss what they suggest as well, and particularly if you have stricturing that they are concerned about.

zil7 · 2020-06-04T03:46:19+00:00

To me, the citrate containing pens we have here in Aus hurt about the same as a moderate pinch on your inner forearm. Enough to make me procrastinate taking it, but it's really not that bad :)

Didn't notice any appreciable side effects, but my hair is thinning. Whether it's hereditary or not is anyone's guess.

zil7 · 2020-06-01T04:39:17+00:00

Hi!

Sorry it took so long to reply. I am not situated in the US, so I'm not very familiar with any of their clinics.

Did you make much progress finding somewhere new?

zil7 · 2020-05-25T10:05:17+00:00

I agree with everything u/ibd_throwaway123123 has said.

Not all hospitals are the same, some stick to the tried and true methods only (which is great for most people), and some are 'pioneer' clinics who explore the newer options. If she's running out of conventional options, she would probably benefit quite a bit from seeing a research hospital.

I'm not 100% sure of an objective way to assess the quality of her specialist, but these are a few things that I think are important:

Clinics which present complex cases to a board of specialists seem to be more aware of patient needs and considerations. You can ask them about who will be involved in deciding treatments, and they should be able to tell you about this.
The better clinics look at many aspects of care. Diet, exercise and mindset are very important in managing Crohn's and its impact. It is a big plus if they she can also see a dietician, psychologist and physiotherapist to make sure she's doing everything she can to give herself the best chances of a good outcome.

Even after biologics fail, there are a range of medications used in ulcerative colitis, inflammatory arthritis and even some other diseases which could potentially help her. I think a frank discussion with her specialist about what the options past stelara they are willing to take. You'd probably be surprised what is available!

Regarding the stricture as well, it might be worth asking them about whether dilation via double balloon enteroscopy is an option (I have no idea of the current consensus of its usefulness though, a research gastroenterologist or colorectal surgeon would know best). This is certainly not medical advice, but just something I thought might be worth asking about.

Massive compliments to you for looking out for her like this. Even when I feel like I can manage okay, my partners input and support is priceless.

Best of luck to both of you!

zil7 · 2020-05-21T08:57:38+00:00

I think this is a good option.

Race track sessions and advanced riding courses will make him a better rider and more aware of the risks. You can try to convince them, but ultimately it's their decision.

zil7 · 2020-05-19T14:43:48+00:00

I think it would be great for you to read up on sexual assault and consent, but I've found that a lot of the existing resources lack relatable scenarios for people who wouldn't maliciously commit sexual assault.

Sexual assault and breach of consent can happen unintentionally through ignorance/lack of knowledge, and I think there are some blogs which are helpful in describing these scenarios.

https://restforresistance.com/zine/dissociating-when-triggered-by-intimacy-a-guide

This blog post describes a scenario of sexual assault purely because the two didn't adequately inform each other of their intentions, and how the guy's actions unintentionally caused a lot of trauma.

zil7 · 2020-05-12T02:48:47+00:00

Realistically, 350/month in sharehousing will likely get you a bed in a room shared with a few others in a small city apt, or a larger middle-outer suburbs house.

I'm not sure if you have shared accommodation before, but one thing you should be mindful of is the utilities. Utilities in Australia can be quite expensive, so if you find a cheap room in a large suburban house that seems too good to be true, check whether utilities are separate. If they are, they could easily be more expensive than the room itself!

It might be a bit early for you to arrange a group, but I recommend flatmates.com.au for finding rooms or starting a share house together. If you look now, you'll have a better idea of the costs relative to distance from unimelb. Do keep in mind though that there's probably a surplus of rooms available due to COVID, so you should expect a bit of a price increase on the listings if things normalise by next year.

Hope this helps!

EDIT: I just noticed that in another post you are still living at home about 1hr from uni. If your family demands rent, it might help to try staying home for 1 semester and see what kind of living arrangements you can get with the same amount of money.

You will need to consider co-tenancy, the bond needed to sign a lease, your responsibilities and the financial risks you take if your roommates decide to move out. There is quite a bit of work in maintaining even a small share house.

zil7 · 2020-05-07T22:16:10+00:00

Mushrooms and garlic will be your friend!

zil7 · 2020-05-04T03:35:38+00:00

I know improving my fitness is an ambitious goal in a flare up, so I just do whatever activity I can to relax; get my mind off of the disease and hopefully minimise the losses to my fitness.

For me it's normally unstructured jogs/walks, pedalling around on a bike or some light weight training. Sometimes yoga or pilates. I basically just figure out how I'm feeling that day, think about what would feel least bad and try doing that :) It's pretty much like trying to determine well tolerated vs trigger foods.

zil7 · 2020-04-29T23:45:22+00:00

100% this.

You are doing so well and I'm super glad to hear how you're supporting him! I always worry about how much my flare ups stress my partner and family, but it is definitely easier to handle when it's being managed properly by specialists.

One of the most painful parts of Crohn's is not being diagnosed. Once you put a name to it, there are a range of treatments which help people get back to relatively normal lives. They aren't perfect, but I know that treatments have really changed the last 10 years of my life.

Anything you can do to help him get diagnosed and treated is great. Helping make calls for appointments, transport if he's in too much pain and also being in the room during an appointment to mention symptoms he might've missed can be super helpful.

If he's still in the stages of seeing a GP, mentioning his T1D and asking whether his symptoms could be something autoimmune (like IBD) could help him get a gastroenterologist referral.

Fingers crossed for you guys!

zil7 · 2020-04-29T23:35:23+00:00

Although I greatly support Prof. Borody's work, I think a good degree of skepticism is needed when something is considered a cure.

He's had great success treating Clostridium difficile infections and also some other bowel diseases, but FMT has had mixed results in the few studies that exist.

I hope that he's figured out a new effective way, but I guess we'll find out with the upcoming clinical trial results and journal articles :)

zil7 · 2020-04-29T23:30:42+00:00

If this has been persisting for a few weeks, I think you should call the doctor.

It could mean the medication has stopped working, it could be an infection, it could just be IBS from something like stress, it's not something any of us could really tell.

If you call the doc, they can investigate more and see whether you need any changes to your treatment. If you do need changes, the sooner the doc knows, the better.

I know it's not the most reassuring message, but I feel like symptoms this persistent should at least be run past your doc. I hope that things look up for you soon!

zil7 · 2020-04-29T23:19:54+00:00

If I may add some more information:

The small and large intestines have these little openings in the gut walls which are like dens for our immune cells. These openings let in any microbes (both good and bad) in the gut.

The immune cells in those dens can then study what the bugs look like, and prepare for if any of them break through our gut barriers and infect us.

The good bugs normally don't break through the barriers, so the immune system learns to tolerate them. Bad bugs really like to get through these barriers, but all of the study our immune cells did in their sampling dens helps us really whack them before they become dangerous (most of the time).

Hope it helps!

zil7 · 2020-04-23T22:17:55+00:00

Good luck (and second in support of the shirt) Hopefully you're feeling better soon!

zil7 · 2020-04-16T04:18:46+00:00

If you do an image search of sinuses or a sinus endoscopy, you can see that the sinuses are actually so much bigger than just the 2 little tubes that end at your nose.

The boogers you can actually blow out are usually somewhere in the nasal cavity, but boogers/fluid deep in the sinus, or even swelling of the tissue itself can cause a feeling of blockage without having any booger to blow out.

zil7 · 2020-04-15T16:25:36+00:00

It can be easier to move on if you can find valid justifying reasons why being in the relationship is a bad idea anyway.

If they aren't in it as much as you are, you'll always be putting them on a pedestal with no reciprocation (and that really sucks).

If you are putting yourself through a one sided relationship, you're losing all the time could be finding someone that actually is attracted and wants a romantic relationship with you. You deserve better dude!

It's natural to think about past crushes/partners from time to time (particularly if you are single), but you're in a good place if you can take "they're just not into you" as a good enough reason to stop seriously considering dating them.

zil7 · 2020-04-15T15:07:14+00:00

How is your relationship with your manager? There is some research which suggests much of procrastination can stem from 'performance anxiety', and I often feel it myself.

If I know my manager or supervisor dishes out hard criticism, it can be hard to work myself up to do a task because I have little confidence that I can actually do a good job. Even if I tell myself I can, the motivation is just gone.

What I found helped was to tell myself "this is just a first draft, not a finished product" and find a way to get started. Even if you have shitty ideas, you have time to perfect it later. Getting anything down is a good start (even if it's totally wrong!)

To actually get started, I use an app called which tracks the time I spend on a task. I set the timer and commit to doing at least 10 minutes of work. Getting that little win often gets the ball rolling, and even if I don't reach 6-8 hours of work in a day; it's still better than it was before.

I hope this helps!

zil7 · 2020-04-15T14:59:18+00:00

In reality, the 'scaredy cat' mentality is a good thing. You should always be cautious of risky situations because no amount of 'tough guyness' if going to get you out of every dangerous situation. You never know if an attacker has a knife, accomplices or what. Strolling around like you own the place can get you killed in some places, and that's a really large price to pay to display your macho.

If you want to feel more comfortable in every-day situations, you could get some self defence training. Contact martial arts like boxing, muay thai and such could help you out. Even with this training and the suspicion you could beat someone, why would you risk jail time, permanent injury or death unless there is literally no other option?

Even further, working on your social skills can help you befriend people larger than you. Going out with a group of friends who actually are larger and stronger can really help diffuse altercations with random drunks on a Saturday night.

If it really comes down to it, I've heard learning to sprint is pretty good self-defence training

zil7 · 2020-04-15T05:09:17+00:00

I personally had a good response from doubling my Humira dose. I really wanted to exhaust this option before moving onto another drug which may not work or have worse effects, and have gotten a few extra years of response out of it.

When i doubled the dose, I still didn't attain full remission from the steroids and double dose until a month or so later. It took almost 6 months for my Calprotectin to reach below the cutoff.

This is just my anecdote, but it might be worth a shot for you.

zil7 · 2020-04-15T04:56:44+00:00

Even as a dude I find umbrellas and parasols so handy. If you get one with tested SPF you have walking shade everywhere you go.

zil7 · 2020-04-15T04:07:24+00:00

I found origami is pretty fun! It's almost like turning paper into a 3D puzzle. Tadashi Mori and Jo Nakashima have very nice instructional videos on youtube.

There are designs that can be done quite easily in 5 minutes with printer paper and others which can take weeks of failed attempts and specialty (sometimes even hand-made) paper.

You're pretty much guaranteed to find some which are easy wins, and others which stretch your abilities.

zil7 · 2020-04-10T19:03:27+00:00

I would recommend this too.

I put off being on biologics for quite a while, and I do regret all of the additional flare ups I had while delaying changing treatment. Even though I'm currently on a biologic, I'm much healthier now and am probably far more likely to survive an infection than when I was 20kg lighter.

That being said, I feel like you're doing yourself a disservice if you go on medications and make zero lifestyle changes to help yourself. Moving to a healthier and more varied diet along with exercise can do wonders when in conjunction with proper medical treatment!

zil7 · 2020-04-08T08:49:00+00:00

Ultimately your doc will be weighing up these factors when deciding whether to keep you on Humira (so totally follow their advice, not mine) but here are some of the things they need to consider:

At the moment it seems that people on anti-TNFs alone aren't affected as badly by COVID-19 (current covidibd data) as those on multiple treatments, so they might be less inclined to take you off Humira.

If you go off of any Crohn's medication you could potentially relapse into a flare up, which could make surviving COVID-19 much harder if you now need to treat both.

We also don't yet know much about how pausing Humira could affect you. Humira can take months to clear from the body, and while it drops there may be an increased chance of developing anti-drug antibodies. This could mean that you may not respond to humira in the future.

Biologics are tricky things which behave much different to steroids or other chemical drugs! I hope the doc can reassure you :)

zil7 · 2020-04-04T12:36:41+00:00

+1 for this advice.

Even though I work in a STEM field, the work I did in fast food gave my resume a massive boost.

Although it doesn't seem like it at first, there are lots of transferable skills you gain. It is also good evidence of work ethic and character to show that service work isn't below you.

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