Verification TH-C-AD;TH-D-DBD
on Monday, September 11th, 2023 9:52 | by Luisa Guyton
![](https://lab.brembs.net/wp/wp-content/uploads/2023/09/Abbildung-8.jpg)
Category: Anatomy, DAN, genetics, Kenyon cells, Optogenetics | No Comments
Cloning via DNA Assembly
on Friday, September 1st, 2023 7:26 | by Isabel Stark
![](https://lab.brembs.net/wp/wp-content/uploads/2023/08/6_16_2023-cPCR-pCFD6-rsh.jpg)
DNA Assembly in a 1:2 ratio of vector to insert with gRNAs of rsh and rut (Q5 and template concentration: 640 pg/µl) and 100ng of pCFD6 BbsI AP (using QuickCIP). Heat-shock (hs) transformation into E. coli (DH5α competent) with 10 µl Assembly Reaction and 100 µl cells.
For Crtl, pCFD6 BbsI AP was wrongly used.
![](https://lab.brembs.net/wp/wp-content/uploads/2023/09/6_26_2023-cPCR-rsh.jpg)
![](https://lab.brembs.net/wp/wp-content/uploads/2023/09/6_29_2023-cPCR-rsh.jpg)
![](https://lab.brembs.net/wp/wp-content/uploads/2023/09/6_29_2023-cPCR-rut.jpg)
DNA Assembly in a 1:2 ratio of vector to insert with the gRNAs of rsh and rut (Q5 and template concentration: 640 pg/µl) and 100ng of pCFD6 BbsI AP (using FastAP and 2 extraction steps). Heat-shock (hs) transformation into E. coli (DH5α competent) with 10 µl Assembly Reaction and 100 µl cells.
For Crtl, pCFD6 BbsI AP was used in reaction.
Category: genetics, Memory, Operant learning, operant self-learning, Radish | No Comments
Line verification SS56699
on Tuesday, August 29th, 2023 7:50 | by Luisa Guyton
After dissecting brains from the GAL4 driver line SS56699 with GFP staining and finding no fluorescence, I dissected them again and the immunohistochemical staining showed fluorescence. To check that the correct neurons were stained, I compared the images with the image in the paper by Hulse et al (2021; https://doi.org/10.7554/eLife.66039) . The three PPL1 dopaminergic dorsal fan-shaped body tangential neurons per hemisphere are stained, but there are some additional unidentified neurons (presumably PPM1 neruons) visible.
![](https://lab.brembs.net/wp/wp-content/uploads/2023/08/image-1.png)
![](https://lab.brembs.net/wp/wp-content/uploads/2023/08/image-4.png)
![](https://lab.brembs.net/wp/wp-content/uploads/2023/08/image-3.png)
![](https://lab.brembs.net/wp/wp-content/uploads/2023/08/image-5.png)
Hulse, B. K., Haberkern, H., Franconville, R., Turner-Evans, D., Takemura, S.-y., Wolff, T., Noorman, M., Dreher, M., Dan, C., Parekh, R., Hermundstad, A. M., Rubin, G. M., & Jayaraman, V. (2021). A connectome of the Drosophila central complex reveals network motifs suitable for flexible navigation and context-dependent action selection. eLife, 10, e66039. https://doi.org/10.7554/eLife.66039
Category: Anatomy, genetics | No Comments
Success: rsh Stock has rsh1 Mutation
on Monday, August 7th, 2023 11:11 | by Isabel Stark
Via gDNA analysis and PCR was the specific area of the rsh gene extracted and amplified where the nucleotide substitution: C to T (Folkers et al., 2006) should be for the rsh1 mutation. The amplicon was Sanger sequenced which proved the nucleotide substitution.
![](https://lab.brembs.net/wp/wp-content/uploads/2023/08/rsh1-mutation.png)
Category: genetics, Memory, Operant learning, operant self-learning, Radish, Uncategorized | No Comments
Creating gRNAs via PCR
on Friday, July 7th, 2023 6:38 | by Isabel Stark
![](https://lab.brembs.net/wp/wp-content/uploads/2023/07/6_15_2023-PCR-rsh-bearbeitet-1024x648.jpg)
![](https://lab.brembs.net/wp/wp-content/uploads/2023/07/6_23_2023-PCR-rut-bearbeitet-1024x266.jpg)
The template pCFD6 was used with a concentration of 640 pg/µl.
![](https://lab.brembs.net/wp/wp-content/uploads/2023/07/6_30_2023-PCR-rsh-bearbeitet-1024x291.jpg)
![](https://lab.brembs.net/wp/wp-content/uploads/2023/07/6_30_2023-PCR-rut-bearbeitet-1024x287.jpg)
The template pCFD6 was used with a concentration of 64 pg/µl (1:10 dilution).
![](https://lab.brembs.net/wp/wp-content/uploads/2023/07/7_6_2023-PCR-rsh-1024x441.jpg)
![](https://lab.brembs.net/wp/wp-content/uploads/2023/07/7_6_2023-PCR-rut-1024x460.jpg)
The template pCFD6 was used with a concentration of 64 pg/µl (1:10 dilution).
50µl of 5xQ5 High GC Enhancer was added to the PCR mix.
![](https://lab.brembs.net/wp/wp-content/uploads/2023/07/7_7_2023-PCR-1-rsh-rut-bearbeitet-1024x666.jpg)
The template pCFD6 was used with a concentration of 128 pg/µl (1:5 dilution).
50µl of 5xQ5 High GC Enhancer was added to the PCR mix.
![](https://lab.brembs.net/wp/wp-content/uploads/2023/07/7_19_2023-PCR-rsh-rut-Phusion-bearbeitet.jpg)
![](https://lab.brembs.net/wp/wp-content/uploads/2023/07/7_19_2023-PCR-rut-Phusion-bearbeitet-1024x587.jpg)
The template pCFD6 was used with a concentration of 640 pg/µl.
The Phusion DNA Polymerase was used instead of the Q5 High-Fidelity DNA Polymerase.
Category: genetics, Memory, Operant learning, operant self-learning, Radish, Uncategorized | No Comments
Gene switch finally worked
on Monday, September 28th, 2020 12:21 | by Andreas Ehweiner
I retried the gene-switch method with a new solution of RU486. Since it did not worked before I just tried one line. The flies were left for two days on instant food mixed with a sugar solution of RU486. This time I saw normal GFP expression. So I will test all 4 line in parallel after the break.
![](https://lab.brembs.net/wp/wp-content/uploads/2020/09/Gen-switch-CD8GFP-1024x768.jpg)
Category: Anatomy, crosses | No Comments
Testing of possible Gal4-lines
on Monday, September 28th, 2020 12:11 | by Andreas Ehweiner
I finished my cross for the colocalisation of FoxP with the 6 Gal4-lines i ordered.
Two lines show a nice colocalisation and will be tested.
![](https://lab.brembs.net/wp/wp-content/uploads/2020/09/Overlapping-lines.jpg)
One would show some overlap, but a completly diffrent expression pattern than it should have. So i will try this line again from the stock to exclude a mixup of the line.
![](https://lab.brembs.net/wp/wp-content/uploads/2020/09/Error-line-2.jpg)
The next one dont seem to have any overlap, but also the pattern looks not like it should.
![](https://lab.brembs.net/wp/wp-content/uploads/2020/09/Error-line-1.jpg)
This line seems to have coexpression but the pattern also looks a bit odd, this may be due to a general weak signal and a bad dissection.
![](https://lab.brembs.net/wp/wp-content/uploads/2020/09/Fraglich-mit-overlap.jpg)
The final line seems to have no overlap with the FoxP expression.
![](https://lab.brembs.net/wp/wp-content/uploads/2020/09/No-overlap.jpg)
Category: Anatomy, crosses, Foxp | No Comments
Test for hsGal4
on Monday, September 14th, 2020 1:40 | by Andreas Ehweiner
Comparison between Control (elavGal4 x CD8GFP) and hsGla4 x CD8GFP. Hs for 3h, fixation and dissection after 24h.
![](https://lab.brembs.net/wp/wp-content/uploads/2020/09/Hs-vergleich-14.9.jpg)
Category: Anatomy, crosses | No Comments
General behavior of the MBON-2
on Monday, August 31st, 2020 12:54 | by Anders Eriksson
I wanted to expand and look into some general behavior of the mbon-2 flies. Mostly as a complement to the already existing data.
![](https://lab.brembs.net/wp/wp-content/uploads/2020/08/General-behaviour-1024x611.jpg)
Category: genetics, Lab, Memory, Operant learning, operant self-learning, Optomotor response, science | No Comments
Progress Week 29
on Monday, July 20th, 2020 1:38 | by Anders Eriksson
-Introduced Sayani to the wonderful world of Drosophila
-Been doing some DTS coding
-Preparing flies to do optomotor response for Mathias Raß
![](https://lab.brembs.net/wp/wp-content/uploads/2020/07/Untitled-1-1014x1024.jpg)
Category: flight, genetics, Lab, Memory, Optomotor response, R code | No Comments