Salt avoidance under blue and redlight

on Sunday, June 29th, 2025 10:10 | by

N=12

N=8

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Gustatory preference results

on Monday, June 2nd, 2025 2:03 | by

Gustatory preference under redlight

Gustatory preference under bluelight

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Gustatory preference results

on Monday, May 26th, 2025 12:29 | by

Gustatory preference under redlight

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Gustatory preference results

on Sunday, May 18th, 2025 11:45 | by

Gustatory preference under redlight

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Verification TH-C-AD;TH-D-DBD

on Monday, September 11th, 2023 9:52 | by

Confocal images with anti-GFP and anti-Brp staining of TH-C-AD;TH-D-DBD > mCD::GFP fly brains: A: 4 PPM 2 DANs per hemisphere marked with blue circles, three clusters of Kenyon cell bodies per hemisphere marked with white arrows. B: Five visible cell bodies of unidentified neurons marked with a blue circle in the left hemisphere. The neurons project into the lobula plate and the medulla. Strong fluorescence of cell bodies of the Kenyon cells projecting into the Mushroom bodies is visible. C: Unidentified neurons projecting into structures outside of the optic lobes especially the ventral lateral protocerebrum.
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Cloning via DNA Assembly

on Friday, September 1st, 2023 7:26 | by

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.

For Crtl, pCFD6 was wrongly used.

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.

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Line verification SS56699

on Tuesday, August 29th, 2023 7:50 | by

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.

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

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Success: rsh Stock has rsh1 Mutation

on Monday, August 7th, 2023 11:11 | by

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.

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Creating gRNAs via PCR

on Friday, July 7th, 2023 6:38 | by

1% Agarose gel with 100bp marker and PCR1 rsh, PCR2 rsh and PCR3 rsh or PCR1 rut, PCR2 rut and PCR3 rut, respectively.
The template pCFD6 was used with a concentration of 640 pg/µl.
1% Agarose gel with 100bp marker and PCR1 rsh, PCR2 rsh and PCR3 rsh or PCR1 rut, PCR2 rut and PCR3 rut, respectively.
The template pCFD6 was used with a concentration of 64 pg/µl (1:10 dilution).
1% Agarose gel with 100bp marker and PCR1 rsh, PCR2 rsh and PCR3 rsh or PCR1 rut, PCR2 rut and PCR3 rut, respectively.
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.
1% Agarose gel with 100bp marker and PCR1 rsh, PCR1 rut.
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.
1% Agarose gel with 100bp marker and PCR1 rsh, PCR2 rsh and PCR3 rsh or PCR1 rut, PCR2 rut and PCR3 rut, respectively.
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.
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Gene switch finally worked

on Monday, September 28th, 2020 12:21 | by

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.

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