Use of the CRISPRiTest assay requires access to a flow cytometer with the following excitation and emission requirements:
Excitation: 488nm and 561nm
Emission: 530/20nm and 590/20nm
Cellecta also offers the following products that may be useful when running this assay:
dCas9-Only Plasmids
| Cat.# | Description | Quantity | Price |
|---|---|---|---|
| SVKRABC9B-PS | CRISPRi dCas9-KRAB pRDKCCB-CMV-dCas9-KRAB-2A-Blast (plasmid) | 25 µg | $300 |
| SVKRABC9H-PS | CRISPRi dCas9-KRAB pRDKCCB-CMV-dCas9-KRAB-2A-Hygro (plasmid) | 25 µg | $300 |
| SVKRABC9B-VS | CRISPRi dCas9-KRAB pRDKCCB-CMV-dCas9-KRAB-2A-Blast (virus) | 1 × 106 TU | $500 |
| SVKRABC9H-VS | CRISPRi dCas9-KRAB pRDKCCB-CMV-dCas9-KRAB-2A-Hygro (virus) | 1 × 106 TU | $500 |
Cell Line
Parental dCas9-KRAB+ MDA-MB-231 Cells (Cat.# ZMDAMB231-CKRA, 106 cells) may be run in parallel with the user’s dCas9-repressor (dCas9-Rep) cells as a positive control for the assay.
Transduction Control Virus
Non-Targeting CRISPR Control pRSG16-U6-sg-UbiC-TagRFP-2A-Puro (virus) may be used to assess transduction efficiency of the target cells.
| Cat.# | Description | Quantity | Price |
|---|---|---|---|
| SGCTL-NT-pRSG16-V | Non-Targeting CRISPR control pRSG16-U6-sg-UbiC-TagRFP-2A-Puro Vector (virus) |
1 × 107 TU | $500 |
Other than the specific reagents and instruments above, the protocol assumes the user has access to standard materials (e.g., polypropylene tubes, pipette tips), equipment (table top centrifuges, pipettes, scales), and common reagents (e.g., TE buffer, ethanol) and buffers used in a typical life science laboratory.
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