跳转至内容
Merck
CN

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.

材料
货号
品牌
产品描述

Sigma-Aldrich
乙酸, glacial, ACS reagent, ≥99.7%
Sigma-Aldrich
氯化钾, ACS reagent, 99.0-100.5%
Sigma-Aldrich
碳酸氢钠, powder, BioReagent, for molecular biology, suitable for cell culture, suitable for insect cell culture
Sigma-Aldrich
盐酸 溶液, 1.0 N, BioReagent, suitable for cell culture
Sigma-Aldrich
氯化钙 溶液, BioUltra, for molecular biology, ~1 M in H2O
Sigma-Aldrich
焦碳酸二乙酯, 96% (NT)
Sigma-Aldrich
氯化镁 溶液, for molecular biology, 1.00 M±0.01 M
Sigma-Aldrich
碳酸氢钠 溶液, solution (7.5%), sterile-filtered, BioReagent, suitable for cell culture
Sigma-Aldrich
D -(+)-葡萄糖, ≥99.5% (GC), BioXtra
Sigma-Aldrich
磷酸钠 一元, ReagentPlus®, ≥99.0%
Sigma-Aldrich
磷酸二氢钠 溶液, BioUltra, 5 M in H2O
Sigma-Aldrich
葡萄糖酸钾, ≥99% (HPLC)
Sigma-Aldrich
DL-2-氨基-5-膦酰戊酸, solid
Sigma-Aldrich
腺苷 5′-三磷酸 二钠盐 溶液, Crystalline ATP, HPLC purified, aqueous solution for RNA transcription
Sigma-Aldrich
SR-95531, ≥98% (HPLC), powder
Sigma-Aldrich
NBQX 二钠盐 水合物, ≥98% (HPLC)
Sigma-Aldrich
鸟苷5′-三磷酸 钠盐 溶液, HPLC purified, aqueous solution for RNA polymerase transcription
Supelco
JWH-018 5-戊酸β-D-葡萄糖醛酸苷 溶液, 100 μg/mL (Acetonitrile:Water (90:10) (v/v)), certified reference material, ampule of 1 mL