- Protocol for Patch-Seq of Small Interneurons.
Protocol for Patch-Seq of Small Interneurons.
STAR protocols (2020-12-31)
Marcela Lipovsek, Lorcan Browne, Matthew S Grubb
PMID33377040
ABSTRACT
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.
MATERIALS
Product Number
Brand
Product Description
Sigma-Aldrich
Adenosine 5′-triphosphate disodium salt solution, Crystalline ATP, HPLC purified, aqueous solution for RNA transcription
Sigma-Aldrich
Sodium bicarbonate, powder, BioReagent, for molecular biology, suitable for cell culture, suitable for insect cell culture
Sigma-Aldrich
Sodium bicarbonate solution, solution (7.5%), sterile-filtered, BioReagent, suitable for cell culture
Supelco
JWH-018 5-Pentanoic acid beta-D-glucuronide solution, 100 μg/mL (Acetonitrile:Water (90:10) (v/v)), certified reference material, ampule of 1 mL
Sigma-Aldrich
Guanosine 5′-triphosphate sodium salt solution, HPLC purified, aqueous solution for RNA polymerase transcription