What are some example applications of pooled whole-genome sgRNA library screens?

Some examples where sgRNA pooled screens have been applied include investigations in: -Mechanisms associated with ALS and Parkinson's Disease (ALS citation: https://www.ncbi.nlm.nih.gov/pubmed/29507424; Parkinson's citation: https://www.ncbi.nlm.nih.gov/pubmed/29269392) -Oncology—looking at growth and metastasis (citation: https://www.ncbi.nlm.nih.gov/pubmed/25748654) -Responses in immune [...]

Where can I find more information about CRISPR/Cas9-mediated gene knockout?

To learn more about CRISPR/Cas9 and the product solutions that we provide, please visit the following pages: Gene editing learning center: https://www.takarabio.com/learning-centers/gene-function/gene-editing Gene editing product catalog: https://www.takarabio.com/products/gene-function/gene-editing The CRISPR/Cas9 system—a simple, RNA-programmable method to mediate genome editing in [...]

How do I perform a genome-wide CRISPR knockout screen?

Below is a general overview of the process of performing a phenotypic screen using a pooled lentiviral sgRNA library. More details are provided in additional sections on this page and in the user manual for the Guide-it CRISPR Genome-Wide sgRNA Library System. STEP 1: select the phenotypic change you wish to study The change must provide a basis for enrichment, selection, or depletion of edited [...]

What should I do if a portion of the recombinant protein passes through a TALON CellThru column without binding to the column?

If a fraction of the his-tagged protein present in the cell extract fails to adsorb to a TALON CellThru column, several explanations may account for this observation: 1. Some types of recombinant proteins may interact with the proteins embedded in the cell membrane. Also, in crude protein lysates, some cellular compartments may not be completely broken down. When an aliquot of crude lysate is [...]

Why does the eluted protein/wash solution look pink?

A pink color in the eluate may be due to the presence of EDTA or DTT (types of metal ion stripping reagents or chelators) in one of the buffers.Protein purification [...]