Direkt zum Inhalt
Merck

Protocol for Patch-Seq of Small Interneurons.

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

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.

MATERIALIEN
Produktnummer
Marke
Produktbeschreibung

Sigma-Aldrich
Essigsäure, glacial, ACS reagent, ≥99.7%
Sigma-Aldrich
Kaliumchlorid, ACS reagent, 99.0-100.5%
Sigma-Aldrich
Natriumbicarbonat, powder, BioReagent, for molecular biology, suitable for cell culture, suitable for insect cell culture
Sigma-Aldrich
Saccharose, ≥99.5% (GC)
Sigma-Aldrich
Salzsäure -Lösung, 1.0 N, BioReagent, suitable for cell culture
Sigma-Aldrich
Magnesiumchlorid -Lösung, for molecular biology, 1.00 M±0.01 M
Sigma-Aldrich
Diethylpyrocarbonat, 96% (NT)
Sigma-Aldrich
Calciumchlorid -Lösung, BioUltra, for molecular biology, ~1 M in H2O
Sigma-Aldrich
Saccharose, BioUltra, for molecular biology, ≥99.5% (HPLC)
Sigma-Aldrich
Natriumbicarbonat -Lösung, solution (7.5%), sterile-filtered, BioReagent, suitable for cell culture
Sigma-Aldrich
D-(+)-Glukose, ≥99.5% (GC), BioXtra
Sigma-Aldrich
Natriumphosphat, ReagentPlus®, ≥99.0%
Sigma-Aldrich
Natriumphosphat monobasisch -Lösung, BioUltra, 5 M in H2O
Sigma-Aldrich
D-Gluconsäure Kaliumsalz, ≥99% (HPLC)
Sigma-Aldrich
DL-2-Amino-5-phosphono-valeriansäure, solid
Sigma-Aldrich
Adenosin-5′-triphosphat Dinatriumsalz -Lösung, Crystalline ATP, HPLC purified, aqueous solution for RNA transcription
Sigma-Aldrich
SR-95531, ≥98% (HPLC), powder
Sigma-Aldrich
NBQX Dinatriumsalz Hydrat, ≥98% (HPLC)
Sigma-Aldrich
Guanosin-5′-triphosphat Natriumsalz -Lösung, HPLC purified, aqueous solution for RNA polymerase transcription
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