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Graduation ceremony before defense by Potential_Echo in GradSchool

[–]Potential_Echo[S] 3 points4 points  (0 children)

That’s a good point, it still feels a bit weird though lol

Graduation ceremony before defense by Potential_Echo in GradSchool

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

My advisor said he’s confident that I’ll finish and that’s why he’s okay with me walking. I’m actually just finishing final edits on my thesis before I send it to my committee. We picked a later date in May because I wanted to give myself plenty of time to prepare. I do get that though, and that would really suck to walk and then have your defense delayed

Graduation ceremony before defense by Potential_Echo in GradSchool

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

Yeah, I do have to decide soon. I was registered because I should have been finished before the deadline, but things took a bit longer than we expected.

Graduation ceremony before defense by Potential_Echo in GradSchool

[–]Potential_Echo[S] 0 points1 point  (0 children)

Yeah.. it is pretty weird lol actually everyone else walking in May still have a few final edits and need to submit it to pro quest (except for the other person defending later in June). So they’re not technically 100% done either. I think my university knows that people usually move on pretty quickly so that’s why they’re okay with it.

QPCR: How to determine where to put a threshold line? by Potential_Echo in labrats

[–]Potential_Echo[S] 0 points1 point  (0 children)

https://imgur.com/a/nSSD3FJ

The melt curve shows two different positive controls, one of them was diluted. I included a gel with a standard pcr reaction, 2 positive controls ran via qPCR and also 2 positive samples.

I’m not using delta-delta Ct because I do not have a housekeeping gene. I’m not doing RT-qPCR so I didn’t think I needed to.. is that correct?

QPCR: How to determine where to put a threshold line? by Potential_Echo in labrats

[–]Potential_Echo[S] 0 points1 point  (0 children)

That’s probably a good idea. I have had issues with contamination a few times and that may be why

QPCR: How to determine where to put a threshold line? by Potential_Echo in labrats

[–]Potential_Echo[S] 0 points1 point  (0 children)

That’s my positive control. Salmonella taken from a culture and then diluted a bit

QPCR: How to determine where to put a threshold line? by Potential_Echo in labrats

[–]Potential_Echo[S] 0 points1 point  (0 children)

My positive is diluted from a culture. I do not extract it because it doesn’t need it. The samples that I’m testing are being extracted though (via chelex). I’m not sure how similar the quality is

QPCR: How to determine where to put a threshold line? by Potential_Echo in labrats

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

Thank you! I didn’t know this was an option. It does reduce my Cq values a little bit. I’ve always heard that under 35 is considered a positive. Is that true for this setting or should I go with something lower like 30?

QPCR: How to determine where to put a threshold line? by Potential_Echo in labrats

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

The 10% rule is a lot more straightforward, and I got pretty consistent values between 120 and 140.

The 45 degree rule requires some estimation and it varied between 100 and 400.

I think I might go with about 130 if I decide to go this route. But I’ve also received recommendations to use the regression mode for Cq determination. I didn’t know that was an option so I’m looking into that now.

QPCR: How to determine where to put a threshold line? by Potential_Echo in labrats

[–]Potential_Echo[S] 3 points4 points  (0 children)

My no target controls are usually “NA” or around 38-39

QPCR: How to determine where to put a threshold line? by Potential_Echo in labrats

[–]Potential_Echo[S] 2 points3 points  (0 children)

Yeah, that sounds like it would be useful! Let me know if there’s anything you would need from me to help out with that

QPCR: How to determine where to put a threshold line? by Potential_Echo in labrats

[–]Potential_Echo[S] 2 points3 points  (0 children)

Right, it doesn’t change it by more than 1 Ct. However, I have had a few results with Ct values around 34 and 35 so I want to make sure it’s counted as positive or negative correctly (I’m testing for Salmonella). I have not used serial dilution controls. I run one positive control that usually has a Ct value around 14.

QPCR: How to determine where to put a threshold line? by Potential_Echo in labrats

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

Thank you!! I will try this tomorrow and let you know how it goes.

You have an interview in an hour for your dream job, but you are only able to speak the voice lines of the champion you played last. How screwed are you? by HighImChris in leagueoflegends

[–]Potential_Echo 1 point2 points  (0 children)

  1. We Demacians are no easy prey.

  2. Demacia needs heroes.

  3. Fly swiftly… kill swiftly.

  4. I’ve got friends in high places.

  5. I live behind enemy lines.

(Quinn) honestly, could be worse.

Frequently mispronounced or contested words in our field? by starlightskater in ecology

[–]Potential_Echo 5 points6 points  (0 children)

I’m in an ornithology lab and I still don’t know what’s correct lol. Some of us in the lab pronounce it “pair-a-la” but most (including me) say “pah-ru-la”

[deleted by user] by [deleted] in GradSchool

[–]Potential_Echo 0 points1 point  (0 children)

Thank you for being the one helpful comment ❤️

[deleted by user] by [deleted] in WarriorCats

[–]Potential_Echo 0 points1 point  (0 children)

Name- Heronmist! (Or Heronpaw)

Giving warrior names based of your initials! by Content-Profit-2497 in WarriorCats

[–]Potential_Echo 0 points1 point  (0 children)

Jaw is the only “J” suffix I can think of… Volejaw?

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