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Determining assembly efficiency by gel electrophoresis, related to step <t>7</t> <t>sgRNA</t> library insert (∼140 bp), digested pLentiCRISPR_v2 backbone (∼13,000 bp), and HiFi assembly reaction product were resolved on a 2% TAE agarose gel. Electrophoresis was performed at 120 V for 30 min in 1× TAE buffer, followed by SYBR green staining and UV visualization. 1 Kb <t>DNA</t> ladder were included for size validation.
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Determining assembly efficiency by gel electrophoresis, related to step 7 sgRNA library insert (∼140 bp), digested pLentiCRISPR_v2 backbone (∼13,000 bp), and HiFi assembly reaction product were resolved on a 2% TAE agarose gel. Electrophoresis was performed at 120 V for 30 min in 1× TAE buffer, followed by SYBR green staining and UV visualization. 1 Kb DNA ladder were included for size validation.

Journal: STAR Protocols

Article Title: In vivo CRISPR screening protocol to identify metastasis mediators using iteratively selected mouse models

doi: 10.1016/j.xpro.2025.104042

Figure Lengend Snippet: Determining assembly efficiency by gel electrophoresis, related to step 7 sgRNA library insert (∼140 bp), digested pLentiCRISPR_v2 backbone (∼13,000 bp), and HiFi assembly reaction product were resolved on a 2% TAE agarose gel. Electrophoresis was performed at 120 V for 30 min in 1× TAE buffer, followed by SYBR green staining and UV visualization. 1 Kb DNA ladder were included for size validation.

Article Snippet: Synthesize the pooled sgRNA oligo library as a custom DNA array using a commercial high-throughput synthesis service (for instance, GeneWiz or GenScript).

Techniques: Nucleic Acid Electrophoresis, Agarose Gel Electrophoresis, SYBR Green Assay, Staining, Biomarker Discovery