The Guide-it CRISPR Genome-Wide sgRNA Library NGS Analysis Kit enables preparation of next-generation sequencing (NGS) libraries from cell populations screened using the Guide-it CRISPR Genome-Wide sgRNA Library System (Cat. # 632646), allowing for identification of sgRNA sequences and corresponding gene knockouts that have increased or decreased in frequency under the conditions of the screen. The kit provides all the components necessary to prepare ten sequencing libraries (20 PCRs), including a genomic DNA purification kit, primers, PCR enzyme and buffers, and a PCR cleanup kit.

The Guide-it CRISPR Genome-Wide Library PCR Kit is designed for NGS library preparation from cell populations screened using the Guide-it CRISPR Genome-Wide sgRNA Library System (Cat. # 632646), and is also sold as part of the Guide-it CRISPR Genome-Wide sgRNA Library NGS Analysis Kit (Cat. # 632647). The kit contains all the primers and reagents necessary for amplification of proviral sgRNA sequences from screened cell populations for analysis by NGS. This kit includes enough reagents for performing 20 PCRs.

Fetal Bovine Serum (FBS) that has been functionally tested for optimal use with all of our Tet Systems. This FBS does not contain trace levels of tetracycline (or its derivatives) which have been observed to interfere with proper regulation of TRE-controlled gene expression. Use of this serum ensures the maximal range of induction possible with the Tet Systems.

Takara's DNA Ligation Kit, Mighty Mix, is a one-solution premix ligation reagent that offers high efficiency ligations, particularly for blunt-ended ligation. The reaction can be performed for 5 minutes at 25°C (rapid protocol), or 30 minutes at 16°C (standard protocol) with equal results for both circular and linear DNA ligations. The 2X Ligation Mix allows small ligation reaction volumes (10 μl) when DNA is limiting. The kit includes sufficient reagent for 100 ligation reactions. Ligation products reaction can be used directly in bacterial transformation.

Stellar Competent Cells are an E. coli HST08 strain that provides high transformation efficiency. These cells can be used in a wide variety of applications—from preparation of cDNA and genomic libraries, to construction of longer-length genomic libraries, to subcloning, and even methylated DNA cloning. Stellar Competent Cells lack the gene cluster for cutting foreign methylated DNA (mrr-hsdRMS-mcrBC and mcrA), and are therefore useful for cloning methylated DNA. The cells can also be used for blue/white screening (i.e., α-complementation) when transformed with vectors containing the lacZα gene. A pUC19 vector is provided as well as SOC Medium. Stellar Competent Cells are recommended for use with Clontech's In-Fusion PCR Cloning Kits.