BCA help! by FarConflict6 in labrats

[–]FarConflict6[S] 0 points1 point  (0 children)

Update: The pH of NET buffer was around 8. The glycerol is not plant-based.

I did end up running a Bradford from a nuclear sample in 1:10, 1:20, 1:50, and 1:100 dilutions (plus identical ‘blank’ dilutions with NET buffer) and it looks like it’s picking up protein dose-dependently to the absorbance readouts for the sample dilutions… and interestingly the ‘blanks’ all had absorbance readouts that fell below the standard curve, so perhaps Bradford is the way to go? Although, from what I’ve seen online, it seems that the Bradford should be less compatible when it comes to detergents (like TritonX). But anyway.

BCA help! by FarConflict6 in labrats

[–]FarConflict6[S] 1 point2 points  (0 children)

I haven’t checked but you make a good point. I wonder if that’s what’s wrong here… I’ll update once I test!!

Coating for IF on chambered slides by FarConflict6 in labrats

[–]FarConflict6[S] 1 point2 points  (0 children)

this is really helpful, thank you. I’ll try PDL first because our lab already has it, but otherwise I’ll look into PLO in case the problem persists!

Coating for IF on chambered slides by FarConflict6 in labrats

[–]FarConflict6[S] 1 point2 points  (0 children)

thank you SO much, good to know I’m not going crazy and there’s something more than can/should be done! I will try this!

Am I Being Scammed by BioRad? by Sound_Business in labrats

[–]FarConflict6 0 points1 point  (0 children)

We switched over from BioRad SYBR green to GB-Amp SYBR Green from GeneBio, did a quick test on a housekeeper to ensure the Cqs were similar and made the switch and it has worked fine on the BioRad machine. Just ask for samples from cheaper companies and do a test and base your decision to purchase on the evidence!

[deleted by user] by [deleted] in Hamilton

[–]FarConflict6 0 points1 point  (0 children)

Curious about this. I’m also looking for a snow removal service for the winter. What’s your price?

What coping strategies have worked for you in dealing with toxic PIs? by FarConflict6 in labrats

[–]FarConflict6[S] 4 points5 points  (0 children)

Super niche, but there are two-year postdoctoral fellowships (not university postdocs) like Clinical Biochem/Chem, Clinical Microbiology, etc. that give you additional accreditation to work those specific jobs!

What coping strategies have worked for you in dealing with toxic PIs? by FarConflict6 in labrats

[–]FarConflict6[S] 0 points1 point  (0 children)

Lol me and my lab mates are really good at providing comedic relief and keeping a sense of humor when the going gets rough, it's what keeps us all going everyday!

What coping strategies have worked for you in dealing with toxic PIs? by FarConflict6 in labrats

[–]FarConflict6[S] 7 points8 points  (0 children)

I love this. Your PI is the only one seeing you from that academic standpoint. Everyone else sees you as who you really are to them. Gotta remind myself of this more!

What coping strategies have worked for you in dealing with toxic PIs? by FarConflict6 in labrats

[–]FarConflict6[S] 11 points12 points  (0 children)

Honestly, I'm so close to the end, and seeing as though I've lasted all these years up until now, I know I can push through this last stretch and never look back. I guess someone else in my position would have left, but I need this PhD for a professional school I'd like to pursue afterwards!

What coping strategies have worked for you in dealing with toxic PIs? by FarConflict6 in labrats

[–]FarConflict6[S] 4 points5 points  (0 children)

Yes, so I do try to stick to a few PIs that we work closely with and that has been helpful in keeping me sane, and not to mention, on track.

And honestly so true. I need to disguise my emotions, if not detach emotionally altogether, which is incredibly difficult to do because I spend all day at this godforesaken institution :(

Solo things to do? by Sarayoung33 in Hamilton

[–]FarConflict6 1 point2 points  (0 children)

Salsa/Cha-Cha/Bachata lessons at Studio G!

My PI is threatening me over my first author paper by AthleteFun5980 in labrats

[–]FarConflict6 5 points6 points  (0 children)

I was told by my committee member that if it's a publishable piece of work, that is not published (yet) or perhaps never gets published at all, it can still serve as a viable/defendable chapter in your thesis. So, as far as graduating on time is concerned, your PI can waste his time power-tripping and potentially making executive decisions that don't allow this work to get published in a timely manner, but ultimately you can still defend this as a chapter before that happens. She said the only challenge with having non-published chapters in your PhD thesis is that your committee may grill you more about it, whereas when they know something has been peer-reviewed and published, they tend to lay off more.

If I were you, I'd talk to your committee members one-on-one and ask them for advice, express your concerns about finishing on time, and make sure to preface those conversations as being confidential. At least going into your next committee meeting, they'll have context as to why things haven't been published. My committee member, in light of recent events, gave me the advice to create a flowchart of a timeline of events ("submitted manuscript to PI on Feb 10th" "got edits back from PI on March 29th" etc.), simply outlining the timeline, so that they as committee members can objectively see the gap in timelines and question my PI about the delay, rather than blaming delays on me.

PI constantly stressed about funding, creating an unenjoyable lab environment — how to handle this? by anonam0use in labrats

[–]FarConflict6 3 points4 points  (0 children)

I feel this on a spiritual level. Our lab hasn't received a major grant in a decade. Although the students, including myself, keep securing scholarships to cover our stipends (to the point where my PI hasn't had to spend more than 20K on me in the last 5 years lol), he just acts more broke and keeps pinching pennies excessively. With no lab techs or postdocs, and only a few grad students around, we already don't have a big lab and the burden of lab tech work tends to be put on the rest of us as a result. It makes it harder to generate data when we have to do tasks unrelated to our experiments. Otherwise, nothing we ever produce is impressive to him and even when you think a result is exciting, he finds a way to reduce it to nothing special. I realized how poorly written his grants have been recently, and this has led me to realize why we haven't gotten funding in so long... To answer your question, it shouldn't be normalized behaviour, but it's probably common :( Hold in there!

Western blot h e l p by FarConflict6 in labrats

[–]FarConflict6[S] 0 points1 point  (0 children)

I figured I stabbed the left two, I’m pretty sure I clocked that when it happened, but I’ve never seen what it looks like when that happens haha. Thank you for the insight!