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Home > 전제품보기 > Genome Editing > Overivew > About Guide-it™ Kits

About Guide-it™ Kits

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Tools for Successful CRISPR/Cas9 Genomic Editing

Use the list below to learn about kits to improve your CRISPR/Cas9 workflow.

In vitro sgRNA production

Use the Guide-it sgRNA In Vitro Transcription Kit to produce high yields of single guide RNAs (sgRNAs) for Cas9/CRISPR studies.

  • Fast, efficient method for sgRNA production—produce any sgRNA in less than 3 hours
  • Transcribe >4 µg of sgRNA per in vitro reaction
  • Resulting sgRNAs can be used for in vitro screening and/or transfection
Guide-it sqRNA In Vitro Transcription Kit workflow

Guide-it sgRNA In Vitro Transcription Kit workflow.

All necessary reagents are included in the kit:

  • Scaffold Template
  • In Vitro Transcription Buffer
  • T7 Polymerase Mix
  • RNase Free Water
  • NucleoSpin Gel and PCR Clean-Up Kit
  • High Yield PCR EcoDry Premix

Testing sgRNA cleavage efficiency

Avoid delivering ineffective single guide RNAs (sgRNAs) to your cells—use the Guide-it sgRNA Screening Kit to test the efficacy of sgRNAs.

  • Highly-optimized in vitro cleavage assay that allows estimation of cleavage efficiency
  • Includes high-quality, recombinant Cas9 protein
Guide-it sgRNA Screening Kit workflow

Guide-it sgRNA Screening Kit workflow.

All necessary reagents are included in the kit:

  • Recombinant Cas9 Nuclease
  • Reaction Buffer
  • 10X BSA
  • RNase Free Water
  • 2 kb Control Fragment
  • Control sgRNA
  • NucleoSpin Gel and PCR Clean-Up Kit
  • High Yield PCR EcoDry Premix

Viral delivery of sgRNA/Cas9

Use the AAVpro CRISPR/Cas9 Helper Free System to deliver expression cassettes for Cas9 and a target-specific sgRNA to hard-to-transfect cells. This system results in significantly higher levels of indel formation than what is typically obtained with plasmid-based delivery.

  • Delivery of Cas9 using AAV precludes genomic integration and persistent expression of Cas9, reducing off-target effects
  • Complete system containing all the reagents necessary to clone the sgRNA and prepare AAV particles
  • Uses a two vector system to overcome the size restrictions of the AAV genome; Cas9 is split between two vectors with a 1.6-kb region of homology that mediates recombination in target cells, producing a full-length Cas9 gene expression cassette
The AAVpro CRISPR/Cas9 Helper Free System workflow

Plasmid delivery of sgRNA/Cas9

The Guide-it CRISPR/Cas9 Systems provide a simple method to generate a vector that will deliver a target-specific sgRNA, Cas9 nuclease, and a bright fluorescent marker to your target cells.

  • A single plasmid expresses all the components necessary for CRISPR/Cas9 targeting of your gene of interest
  • The plasmid also expresses an exceptionally bright fluorescent protein (tdTomato or ZsGreen1) to monitor transfection efficiency and/or enrich for transfected cells by flow cytometry
  • Plasmid construction is simple; design oligos specific to your target sequence, and clone into the pre-linearized plasmid using the included ligation reagents and high-efficiency competent cells
pGuide-it vector

Gesicle delivery of sgRNA/Cas9

Use the Guide-it CRISPR/Cas9 Gesicle Production System to deliver Cas9 protein and a custom sgRNA to a broad range of cell types with very low cytotoxicity.

  • Direct delivery of Cas9 protein instead of plasmid-based delivery prevents overexpression and genomic integration of Cas9, reducing off-target effects.
  • Tight control over the dose and timing of delivery.
  • The user makes gesicles containing active Cas9-sgRNA ribonucleoprotein (RNP) complexes from a producer cell line. Gesicles are harvested from the supernatant and added to the target cells.
Guide-it CRISPR/Cas9 Gesicle Production System

RNA delivery of sgRNA/Cas9

Use the Xfect RNA Transfection Reagent to efficiently transfect mammalian cells with messenger RNA (mRNA) and/or single guide RNA (sgRNA) with very low cytotoxicity.

  • Transfection with Cas9 mRNA prevents genomic integration and persistent expression of Cas9, reducing off-target effects
  • Simple, serum-compatible protocol
  • Cotransfect with DNA using the Xfect Transfection reagent
Guide-it indel identification kit overview

Monitoring the efficiency of genome editing

The Guide-it Mutation Detection Kit provides a simple PCR-based technique to identify insertions and/or deletions generated in mammalian cells using CRISPR/Cas9 technology.

  • Faster and more efficient than the Surveyor assay—amplify directly from your target cells
  • Complete kit—Guide-it Resolvase, Terra PCR Direct Polymerase, and all buffers are included
Guide-it Mutation Detection Kit workflow

Guide-it Mutation Detection Kit workflow.

All necessary reagents are included in the kit:

  • Cell Extraction Buffers
  • Guide-it Resolvase
  • Terra PCR Direct Polymerase Mix
  • Terra PCR Direct Buffer
  • PCR-Grade Water

Confirming monoallelic or biallelic mutations

The Guide-it Genotype Confirmation Kit provides a simple method to determine if a given clone has mutations in one allele (monoallelic), both alleles (biallelic), or is unchanged (wild type). Expedite the laborious process of screening a large number of clones for those with the desired genotype.

  • Streamlined protocol that uses direct amplification of target cellular genomic DNA
  • Includes highly purified recombinant Cas9 nuclease for the in vitro cleavage reaction
Guide-it indel identification kit overview

Indel identification

The Guide-it Indel Identification Kit provides a complete workflow for identifying the variety of indels created by CRISPR/Cas9 gene editing in a cell population.

  • Kit includes components needed to amplify, clone, and prepare modified target sites for DNA sequence analysis.
  • Workflow has been optimized for fast results.
Guide-it indel identification kit overview