ALM (AP 2.5 mm; ML 1.5mm; diameter 1.5 mm) is the cortical area that produced behavioral effects with photoinhibition during the delay epoch 3 (link),24 (link). For the thalamic reticular nucleus the coordinate was AP -0.7, ML 1.6, DV 3.7 - 3.3 mm, as retrograde labeling from thalALM showed labeling in this sector of the thalamic reticular nucleus (Extended Data Fig. 9 ). Virus and tracer were injected through the thinned skull using a volumetric injection system (modified from Mo-10 Narishige) 48 (link). Glass pipettes (Drummond) were pulled and beveled to a sharp tip (outer diameter ∼ 20 - 30 μm), back-filled with mineral oil and front-loaded with viral suspension immediately before injection. The injection rate was 15 nl/min. See Supplementary Table 2 for description of viruses and injection coordinates used for each experiment. We used the following viruses and tracers: AAV2/1-CAG-EGFP (Penn vector Core, University of Pennsylvania), AAV2/10 CAG-flex-ChR2(H134R)-tdTomato (Penn vector Core, University of Pennsylvania), AAV1-CAG-mRuby2-FLAG49 , WGA-Alexa Fluor® 555 (Thermo Fisher Scientific)50 (link) (WGA555), and Red RetroBeads (Lumafluor).
Aav1 cag mruby2 flag49
Manufactured by Thermo Fisher Scientific
AAV1-CAG-mRuby2-FLAG is a recombinant adeno-associated virus (AAV) vector that expresses the fluorescent protein mRuby2 with a FLAG tag under the control of the CAG promoter. The vector is based on the AAV1 serotype.
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2 protocols using aav1 cag mruby2 flag49
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Targeted neural circuit manipulation
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Targeted neural circuit manipulation
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ALM (AP 2.5 mm; ML 1.5mm; diameter 1.5 mm) is the cortical area that produced behavioral effects with photoinhibition during the delay epoch 3 (link),24 (link). For the thalamic reticular nucleus the coordinate was AP -0.7, ML 1.6, DV 3.7 - 3.3 mm, as retrograde labeling from thalALM showed labeling in this sector of the thalamic reticular nucleus (Extended Data Fig. 9 ). Virus and tracer were injected through the thinned skull using a volumetric injection system (modified from Mo-10 Narishige) 48 (link). Glass pipettes (Drummond) were pulled and beveled to a sharp tip (outer diameter ∼ 20 - 30 μm), back-filled with mineral oil and front-loaded with viral suspension immediately before injection. The injection rate was 15 nl/min. See Supplementary Table 2 for description of viruses and injection coordinates used for each experiment. We used the following viruses and tracers: AAV2/1-CAG-EGFP (Penn vector Core, University of Pennsylvania), AAV2/10 CAG-flex-ChR2(H134R)-tdTomato (Penn vector Core, University of Pennsylvania), AAV1-CAG-mRuby2-FLAG49 , WGA-Alexa Fluor® 555 (Thermo Fisher Scientific)50 (link) (WGA555), and Red RetroBeads (Lumafluor).
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