Sh rna brochure
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Sh rna brochure Document Transcript

  • 1. Guaranteed Knock Down for Every Human, Mouse, and Rat Gene Our experimentally verified shRNA design algorithm ensures the maximum gene-specificity and RNAi efficacy. We guarantee that you will see 70% or greater knock down for your targeted gene with at least two of the four shRNA plasmids.* The availability of two effective sequences allows you to properly control for nonspecific and off-target effects. Enrich or Select: Use Neomycin or Puromycin Resistance for stable selection and study the long term effects of gene suppression. Use GFP Marker for FACS enrichment and the study of short term effects of gene suppression. SureSilencing shRNA Plasmid Kit Contents: 4 shRNAs targeting gene-of-interest 1 negative control shRNA Plasmid Options: For FACS Sorting: GFP Marker (Cat. Nos. KX#####G) For Stable Selection: Neomycin (Cat. Nos. KX#####N) Puromyin (Cat. Nos. KX#####P) Prepare stock of transfection-grade plasmids by amplification in bacteria. Transfect shRNA Plasmids into Cell Line of Interest. Transfect all 4 shRNAs and Negative Control into your cell line in a six-well culture plate. Guaranteed!*: Knock down expression of the target gene by at least 70 percent. Control for non-specific and off target effects. Convenient and Cost-Effective: Use standard plasmid-based and lipid-mediated transfection methods. Plasmids provide a renewable source of RNA Interference. e ativ Neg 4 NA shR 3 NA shR 2 NA shR 1 NA shR hairpin RNA (shRNA) designs packaged in a plasmid backbone. 3 days interference. For each gene, we provide four separate short How It Works shRNA 1 shRNA 2 shRNA 3 shRNA 4 24 - 48 hours the expression of every human, mouse, or rat gene by RNA Negative If using GFP Marker Plasmid, Analyze Short Term Effects of Gene Suppression Enrich by FACS or analyze gene function by fluorescence microscopy. > 1 day SureSilencing™ shRNA Plasmids specifically knock down If using Selection Markers, Analyze Longer Term Effects of Gene Suppression 100 80 Select for stable transfectants using Neomycin or Puromycin. 60 1 - 2 weeks - OR- Guaranteed Knock Down Verify Suppression of Gene Expression Using Real-Time PCR 40 hours Relative Expression Level (Percent of Control) RNA InterFerence SureSilencing™ shRNA Plasmid-Based RNA Interference: 20 0 MAP3K1 ITGB1 MYC SP1 STAT1 HSPCB Targeted Gene SureSilencing™ shRNA Plasmids knock down target gene expression in transfected cells by at least 70 percent. SureSilencing shRNA Plasmids with GFP Marker for each of six genes were transfected into HEK-293 cells. The transfection efficiency was estimated by fluorescence microscopy. The relative expression level of each target gene, normalized to transfection efficiency was determined by qRT-PCR and plotted with the standard deviation of the biological and technical replicates. Perform biochemical, cell, or molecular biological assay of gene function
  • 2. Feature Summary Tested Experimentally verified design algorithm ensures efficacy. Specificity Minimizes off-target effect with the Smith-Waterman specificity search. Plasmid Backbone U1 promoter transcribes moderate shRNA amount: minimal off-target effect & toxicity Bacterial origin of replication and ampicillin-resistance marker: amplify life-time supply Mammalian Expression Markers: Neomycin or Puromycin Resistance: select stably transfected cells , study long term effects GFP Marker: enrich or track transiently transfected cells, study short term effects Unlike chemically synthesized siRNA, plasmid-based shRNA provides a renewable source of RNA interference. You can Applications Use RNAi in virtually any cell line: Accommodates both low and high transfection efficiencies. amplify enough plasmid to complete any number of projects in your lab. Purchase of SureSilencing™ shRNA plasmids includes complete shRNA sequence information. Stable transfections with resistance markers: Study slow effects using any gene function assay. Transient transfections GFP-enriched by FACS: Study quick effects using any gene function assay. * The SureSilencing™ shRNA Guarantee: At least two of the provided pre-designed SureSilencing™ shRNA Plasmids are guaranteed to knock down expression of the targeted gene at the RNA level by at least 70 percent as measured by real-time qRT-PCR in transfected cells upon FACS-based enrichment for GFP expression or selection for neomycin or puromycin resistance as described in the User Manual. Transient transfections GFP-tracked by fluorescence microscopy: Study quick effects using cell-based imaging and gene function assays. SureSilencing™ shRNA Plasmid Ordering Guide Find the shRNA for your genes at: Cat. No. KX#####N KX#####P KX#####G www.SABiosciences.com/RNAisearch.php Description SureSilencing™ shRNA Plasmid with Neomycin Resistance SureSilencing™ shRNA Plasmid with Puromycin Resistance SureSilencing™ shRNA Plasmid with GFP Marker X = H for Human, M for Mouse, or R for Rat Related Products: For Real-Time PCR Verification of Knock Down: RT2 PCR qPCR Assays: Available for all shRNA target genes and housekeeping genes RT2 Real-Time™ SYBR Green Master Mix RT2 First Strand Kit (C-03) Focus on your Pathway SABiosciences Corporation 6951 Executive Way Frederick, MD 21703 www.SABiosciences.com T 301.682.9200 888.503.3187 F 301.682.7300 888.465.9859