- Description Intact Genomics is your resource for in vitro gene editing using Cas9 enzymes. We understand your need to generate sufficient quantities of sgRNA for in vitro gene editing projects. The IG® sgRNA synthesis kits are simple to use, and our kits are scalable to help you reach these goals. The IG® sgRNA synthesis kits can be purchased with or without the sgRNA purification kit. Benefits IG® sgRNA Synthesis Kits are a perfect choice for a variety of sgRNA synthesis needs. Below is a sampling of some of the key benefits. IG® sgRNA Synthesis Kit includes everything but your targeting oligo (all buffers, enzymes, scaffolds) User simply needs to provide the oligo for your target—Must be 55 base pairs and generated using the template design in this manual. Rapid Workflow (Less than 1 hour) Scalable Yield (just 1 reaction for all the sgRNA you need) Customer support – our team is available to aid with your success. The process of sgRNA synthesis by in vitro transcription (IVT) is intricate but allows for successful creation of longer sgRNAs (e.g. 100 or more nt) that cannot be easily chemically synthesized and purified at a low cost. sgRNA synthesis can often be complicated and the maximum yield of sgRNA depends on the oligo target. Despite this complexity, it is important to use sufficient sgRNA to demonstrate the efficiency of in vitro DNA cutting for your gene editing. The IG® sgRNA synthesis kit solves this issue by providing at least 10ug of sgRNA per reaction (100 ul), enough for in vitro DNA cleavage/editing in most cases. Various competitors’ kits can only claim production of a minimum of 4µg of RNA. The increased yield from the IG® sgRNA synthesis kits will enhance your chances of experimental success. This kit provides large sgRNA synthesis volumes to get more yield. For successful sgRNA Synthesis, scientists on your team need simply to identify a DNA modification target of interest and order unique primers targeting that sequence. From there, our team provides a simple workflow to quickly complete sgRNA synthesis, so that you may progress to gene editing using IG® Cas enzymes. Intact Genomics is with you every step of the way to complete your projects. We offer expert advice, fast ordering and delivery, and product customization. Components and Storage IG® sgRNA Synthesis Kits contains the items below. Store all components at -20°C. IG® sgRNA Enzyme Mix (10X) IG® sgRNA Reaction Mix, S. pyogenes (4X) DNase I (RNase-free, 2 Units/uL) IG® sgRNA Control Oligo, S. pyogenes (1 uM) Dithiothreitol (DTT, 0.2 M) NTPs mix (25 mM) Items needed but not provided in this kit: Nuclease Free Water Custom 55-nucleotide oligo (see protocol tab for instructions for designing your oligo) Nuclease-free pipette tips and microcentrifuge tubes Comparative Data We compared the yields of the IG® sgRNA Synthesis Kit for Cas 9 nuclease with the New England BioLabs Inc. EnGen® sgRNA Synthesis Kit and consistently achieved greater yields of gRNA in four experiments. 3203 3206
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