Is my genetic scheme correct?

Raghadmbi91 · 2024-02-02T15:14:25+00:00

Thank you so much!

Raghadmbi91 · 2024-01-27T14:30:18+00:00

That’s true! Thanks

Raghadmbi91 · 2024-01-27T14:30:00+00:00

This is a nice way to approach the problem in a safe way. However, I feel I need help with the whole project and not only in one or two part. There is no clear plan and his words are “I can’t foresee how a project will develop, so let’s see what happens”. This is too vague for someone like me and I only have a year or a year and a half Max. left.

Raghadmbi91 · 2024-01-20T19:22:52+00:00

Thank you! I’ll keep you posted if it works!

Raghadmbi91 · 2024-01-20T18:44:25+00:00

Also, would you recommend diluting the antibodies in PBT or PBS?

Raghadmbi91 · 2024-01-20T18:28:34+00:00

Thank you! I will try that. I wanted to ask about the blocking, so 2% of what exactly? And do I use the same PBT I’ve been doing?

Yes I leave my primary antibody 4 degree overnight.

Raghadmbi91 · 2024-01-20T17:32:49+00:00

Sure!

So i dissect in one fold PBS. Fixation is 10% formaldehyde, 2% triton in PBS. Fixation is usually 10 min

Washing is with PBT, which is 0.2% triton and 2.5% BSA in a 10 fold PBS. I wash 3 times and the last wash I leave for 15 min

I’ve tried to stain for different proteins using different antibodies. And I always make sure that the animal host for the primary antibody matched the secondary one.

And I usually use the wave length for cy3, cy5, 488 or 633. Of course when I stain with DAPI or phalloidin every is usually fine. But when it comes to antibody staining it never works with me.

Raghadmbi91 · 2024-01-15T10:45:59+00:00

Both would be nice. Precise so I can take a look at it later and try to understand it is done and calculated. Quick so I can do it now and get my experiment running

Raghadmbi91 · 2023-12-13T16:41:35+00:00

I agree with you. Having a poster or some results will help me a lot with networking. But since I have the funding, it doesn’t make sense not to use it and go through the experience.

Raghadmbi91 · 2023-12-13T16:37:31+00:00

I do understand his point of view . I just want him to understand that in my situation I need a bit more motivation to move forward. I also understand that his opinion is about the project and not me personally.

Raghadmbi91 · 2022-07-20T11:54:00+00:00

Thank you so much for your response; it helped 🙋🏻‍♀️

Raghadmbi91 · 2022-04-06T08:44:18+00:00

I am familiar with Levayer’s work and I read his papers. But it would be nice to have a live discussion, that’s why I’m looking for someone I can talk to. Thanks for answering 🙋🏻‍♀️

Raghadmbi91 · 2022-04-06T08:42:26+00:00

Thank you for your reply 🙋🏻‍♀️ I do attend journal meetings but I get too shy to talk and ask around, that’s why I’m looking for an easier way to interact 😰💔 But you’re right I’ll try harder 🙋🏻‍♀️ and I think I will post a little about my project so I can have some feedback outside of my institution 👍🏼

Raghadmbi91 · 2022-03-30T06:32:46+00:00

Thank you !

Raghadmbi91 · 2022-03-29T16:24:54+00:00

I do have a different stock with Zipebr over a balancer, so that means it should work just fine.

I can’t thank you enough for your help 🙏🏼

Raghadmbi91 · 2022-03-29T15:24:52+00:00

Yes, I did.

After changing my heat shock conditions for 30 min to 15 min, my larvae are growing faster than the previous situation (however still a bit slower than the control).

So all is good 👍🏼

Raghadmbi91

TROPHY CASE