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1, What is the sgRNA?
Answer: A single guide RNA (sgRNA, also known as guide RNA or gRNA) is a RNA sequence that can guide the Cas9 to the desired location of the cell’s genome.
2, What delivery formats do you provide the sgRNA libraries in?
Answer: Lentiviral particles, transfection-ready DNA, and bacterial stock.
3, Are the sgRNA libraries available as individual clones?
Answer: Yes. Currently, our sgRNA libraries are available standard as pools. However, if you wish to receive your libraries as individually arrayed clones, simply contact us for a custom quote.
4, Can I co-transduce the sgRNA and Cas9 lentiviral particles?
Answer: Yes. However, we strongly recommend that you first establish a cell line which is stably expressing Cas9, in order to best maintain consistent representation of each sgRNA in the library. First, transduce your cells with the Cas9 lentiviral clone. Once you have established the stable Cas9-expressing cell line, use the sgRNA libraries for transduction.
5, Can you ensure that a single cell could be infected with one single sgRNA in the library?
Answer: You can't completely ensure that every cell is infected with one sgRNA, or that all sgRNAs are represented in the pool of infected cells. However, these libraries are small, so if the number of infected cells was 100 × the number of sgRNAs in the library, then you should have close to complete representation.
6, Can the sgRNA lentiviral plasmids be transfected instead of using lentivirus?
Answer: Yes. The lentiviral plasmids are "dual-use", so that they can either be packaged into lentiviral particles or transfected into cells by standard transfection protocols.