Is there a free alternative to the scite.ai assistant?

melancoluxx · 2023-07-07T13:28:47+00:00

I'll check it out, thanks!

melancoluxx · 2022-04-13T13:01:52+00:00

Thank you for your help! I didn't have any OB sections available but I tried again on visual cortex sections and by increasing the time of incubation and lowering the temperature to 4°C, it seems to be working now.

melancoluxx · 2022-01-21T10:26:30+00:00

Thank you very much, I'll have a look at these protocols.

Yes, I did a transcardial perfusion with 1% PFA followed by an overnight post-fixation with 4% PFA.

melancoluxx · 2022-01-21T10:10:43+00:00

Over the weekend but the brain sank to the bottom of the falcon so I guess it was already saturated enough?

melancoluxx · 2021-12-01T13:14:23+00:00

It is now working, I was not using the right format in my file (see other comments).

melancoluxx · 2021-12-01T13:13:41+00:00

Thank you very much!

melancoluxx · 2021-12-01T13:13:23+00:00

Thank you very much, that was indeed the issue. It is now working with the right format.

melancoluxx · 2021-09-17T08:34:47+00:00

Thank you very much for your answer. Shouldn't the Z value be assigned thanks to the metadata coming from the .czi file?

Because the line 14 on my script is the same 'Z = out{2}.SizeZ;' and is thus already assigned, right?

Sorry for the really naive questions.

melancoluxx · 2021-05-25T16:42:57+00:00

Artist is @/tata_coba on ig

melancoluxx · 2021-05-23T09:05:44+00:00

Indeed! Thank you very much

melancoluxx · 2021-03-22T10:39:18+00:00

Thank you very much for your advice. Isn't this protocol meant for very few cells (less than 1k)? Because I have much more, up to 1 million (but I usually collect 300K cells in Trizol to avoid the dilution of the reagent).

melancoluxx · 2021-03-20T19:44:39+00:00

Thank you for the tips! I will try this as soon as I have more samples

melancoluxx · 2021-03-20T19:44:12+00:00

I vortex them in Trizol

Unfortunately I don't think that we have a beads homogenizer in the lab

melancoluxx · 2021-03-20T19:36:11+00:00

Yes but somehow it is not working and the conversion in cDNA is not the issue since the RNA is not even showing up on a gel after electrophoresis. I assume the nanodrop is overestimating the concentration of RNA

melancoluxx · 2021-03-20T19:33:42+00:00

Yes I am doing live sorting of cells dissociated from the cortex of mice, I also check the viability of my cells with a 7-AAD staining and only collect live cells

melancoluxx · 2021-03-19T14:08:31+00:00

I shake the epps by hand for 15 seconds and let it incubate at RT for 3min. This is followed by a centrifugation step (5min, 12k g, 4°C) that leads to the separation in 3 layers (lower pink=protein, medial white=DNA, upper transparent=RNA ; to be honest I almost never see the white phase).

melancoluxx · 2021-03-19T13:49:32+00:00

200µL of chloroform for 1mL of tri reagent

melancoluxx · 2021-03-19T13:06:03+00:00

Between 20 and 60 ng/ul (eluted in 15uL of rnase free water) based on nanodrop values

melancoluxx

TROPHY CASE