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Merck

Protocol for Patch-Seq of Small Interneurons.

STAR protocols (2020-12-31)
Marcela Lipovsek, Lorcan Browne, Matthew S Grubb
摘要

Obtaining electrophysiological recordings and gene expression information from the same neuron (Patch-seq) brings forward a unique opportunity to study the transcriptional correlates of functional properties and vice versa. Here, we provide a detailed Patch-seq protocol tailored to the specialized demands of studying small interneurons. Focusing on the technically demanding process of transitioning between patch recordings and cell extraction, our protocol describes and troubleshoots steps for successfully collecting small interneurons, allowing for multi-modal Patch-seq interrogation of this crucial cell type.

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乙酸, glacial, ACS reagent, ≥99.7%
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氯化钾, ACS reagent, 99.0-100.5%
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碳酸氢钠, powder, BioReagent, for molecular biology, suitable for cell culture, suitable for insect cell culture
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蔗糖, ≥99.5% (GC)
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盐酸 溶液, 1.0 N, BioReagent, suitable for cell culture
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氯化镁 溶液, for molecular biology, 1.00 M±0.01 M
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焦碳酸二乙酯, 96% (NT)
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氯化钙 溶液, BioUltra, for molecular biology, ~1 M in H2O
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蔗糖, BioUltra, for molecular biology, ≥99.5% (HPLC)
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碳酸氢钠 溶液, solution (7.5%), sterile-filtered, BioReagent, suitable for cell culture
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磷酸钠 一元, ReagentPlus®, ≥99.0%
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