Experiments on larvae locomotion using control and experimental line

on Thursday, November 26th, 2020 10:55 | by

Experiment on larvae using the control line (ELAV-Gal4 x UAS-gRNA) and experimental line (ELAV-Gal4 x UAS-gRNA-Cas9).
The experiment was significant, with the p-value p=0.0003
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PCR with undigested pCDF6

on Monday, November 23rd, 2020 12:54 | by

primer: reverse/forward each 10 µl
dNTPs: 4 µl
buffer: 40 µl
polymerase: 2 µl
H2O: 133.6 µl
undigested vector: 0.4 µl

PCR of undigested vector
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Further experiments on larval locomotion using Canton-S wild type flies

on Sunday, November 22nd, 2020 10:33 | by

All experiments were done using 1 mm graph paper and a microscope to count the boxes more precisely.

Tracking of the larvae using Ctrax

Agar plate dyed with activated charcoal (2 g charcoal/ 1 l H2O)
Tracking with Ctrax software using the Burridan camera setup.
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Dprime and D1 were verified

on Monday, November 16th, 2020 11:14 | by

Dprime
D1
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Local FoxP knock out

GMR65A06
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Experiments on larval locomotion using Canton-S wild type flies

on Friday, November 13th, 2020 11:07 | by

Time Experiments

One larva with a time exceeding 20 minutes was not considered.
Two larvae with a time exceeding 20 minutes were not considered.

Distance Experiments

Experiment 4: Graph Paper

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Set-Up test

on Monday, November 9th, 2020 11:03 | by

Wtb
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pCDF6 cloning with three inserts

on Sunday, November 8th, 2020 7:33 | by

pCDF6 digestion with enzyme Bbs1
-5.5µl vector (undigested), conc.: 539.1 ng/µl
-1 µl enzyme (Bbs1)
-5 µl buffer (CutSmart)
-38.5 µl H2O
Agarose-gel electrophoresis
E.Z.N.A. Gel Extraction-DNA Purification from Agarose gel

7Concentration: 38,0 ng/µl
pCDF digested
concentration: 38,0 ng/µl

PCR pCDF6 Primer
-12.5 µl primer reverse/forward
-5 µl dNTPs
-2.5 µl Taq-Polymerase
-0.29 µl pCDF6 digested
-50 µl buffer
-167.5 µl H2O
Agarose-gel electrophoresis
E.Z.N.A. Gel Extraction

Concetration:
primer 1: 170.7 ng/µl
primer 2: 137.4 ng/µl
primer 3: 215.0 ng/µl
primer with 100kb ladder

pCDF6 NEBuilder Assembly Reaction
-pcr1: 4.85 ng -> 1.60 µl
-pcr2: 4.24 ng -> 1.20 µl
-pcr3: 4.87 ng -> 1.20 µl
-pCDF6: 100 ng -> 2.90 µl
-H2O: 3.1 µl
-10 µl NEBuilder HiFi DNA Assembly Master Mix/Control

heat shock transformation
-> plated on LB0+Amp plates
-> no colonies

colony PCR (29.10.20)
-37 µl primer forward/reverse
-37 µl dNTPs
-37 µl Taq Polymerase
-74 µl buffer LSB
-518 µl H2O
Agarose-gel electrophoresis


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New evaluation of the Rover/Sitter larvae

on Thursday, November 5th, 2020 12:34 | by

Control Group 1 has a higher density of larvae compared to Control Group 2.

Percentage of Sitter and Rover larvae, who stayed on the first food patch after 20 minutes.

Course Evaluation

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Gen-switch lines

on Monday, November 2nd, 2020 11:51 | by

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