Use of the CRISPRaTest 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 |
|---|---|---|
| SVVPHC9B-PS | CRISPRa dCas9-VPH pRDVCCB-CMV-dCas9-VPH-2A-Blast (plasmid) | 25 µg |
| SVVPHC9B-VS | CRISPRa dCas9-VPH pRDVCCB-CMV-dCas9-VPH-2A-Blast (virus) | 1 × 106 TU |
Additional dCas9-VPH plasmids are available. Please visit our CRISPRa dCas9-VPH Expression Construct web page.
Cell Line
dCas9-VPH+ MDA-MB-231 Cells (Cat.# ZMDAMB231-CVPH, 106 cells) may be run in parallel with the user’s dCas9-transactivator (dCas9-TA) 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 |
|---|---|---|
| SGCTL-NT-pRSG16-V | Non-Targeting CRISPR control pRSG16-U6-(sg)-UbiC-TagRFP-Puro Vector (virus) |
1 × 107 TU |
Non-targeting controls in other Cellecta vectors are available. Please visit our CRISPR Non-Targeting sgRNA Control Construct in Standard Lentiviral Vector web page.
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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