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Protocols and Applications

Protocol

Enrichment of transfected cells with Dharmacon Edit-R Fluorescent dCas9-VPR mRNA

Edit-R Fluorescent dCas9-VPR mRNA enables both transfection optimization and enrichment for gene modulation experiments. For optimal enrichment of edited cells, sorting the cells for high EGFP expression levels (selecting the top 10% fluorescence) in addition to negative and dim fluorescence for com... More


Protocol

Dharmacon Edit-R Lentiviral sgRNA glycerol stocks

The Dharmacon™ Edit-R™ Lentiviral sgRNA vector is part of the Edit-R CRISPR-Cas9 system for genome engineering. The purpose is to provide the researcher with the most effective tools to deliver a gene-specific sgRNA and, together with Cas9 expression, allow gene editing in cells. In addition to the ... More


Protocol

CRISPRmod dCas9-VPR or dCas9- SALL1-SDS3 mRNA and synthetic guide RNA transfection protocol

The following is a protocol for transfecting CRISPRmod dCas9-VPR or dCas9-SALL1-SDS3 mRNA with synthetic guide RNA into cultured mammalian cells using DharmaFECT™ Duo transfection reagent (Cat #T-2010-xx). The protocol is written for transfection into 96-well tissue culture plates.... More


Protocol

IsoCode Single-Cell Innate Immune: Human CD34+ Myeloid Stem Cell Protocol

A. Overview1. Overview of ProtocolDay 1: Cryopreserved cells are thawed and cultured overnight in the presence of IL-2.Day 2: Enrichment and Stimulation of CD34+ cells for 24 hours.Day 3: Staining and Loading of CD34+ cells onto IsoCode chip.NOTE: This protocol outlines the standard method for thawi... More


Protocol

IsoCode Single-Cell Innate Immune: Human Monocyte Protocol Using Membrane Stain

A. OverviewOverview of ProtocolDay 1: Cryopreserved cells are thawed and cultured overnight in complete RPMI media.Day 2: Enrichment and Stimulation of Monocytes for 24 hours.Day 3: Staining and Loading of Monocytes onto IsoCode Chip.... More