...Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors gRNAs...Figure 1: Overview of the basic CAST mechanism. Created with BioRender.com. Browse, sort, or search the...

...Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors gRNAs... in the DNA. Indels often lead to frameshifts, creating loss of function alleles. To introduce specific...

...Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors gRNAs... in the DNA. Indels often lead to frameshifts, creating loss of function alleles. To introduce specific...

...AmpR 26095 pFHMSP-LIC-N N/A Expression vector for secreted proteins with N-terminal hexahistidine tag and...AmpR 26100 pFHMSP-LIC-C N/A Expression vector for secreted proteins with C-terminal hexahistidine tag and...

...paper . How to Cite this Kit These plasmids were created by your colleagues. Please acknowledge the Principal...Investigator, cite the article in which they were created, and include Addgene in the Materials and Methods...

...help to increase the density of the culture. Normally cultures shake at 150 - 250 rpm, increase this to...Streaking and Isolating Bacteria on an LB Agar Plate Creating Bacterial Glycerol Stocks for Long-term Storage...Autoclave and allow to cool to room temperature. Now screw on the top of the bottle and store the LB at room... storage of the bacteria, you can proceed with Creating a Glycerol Stock . You can now isolate your plasmid...

...collection of Addgene's video content, including how-to screen captures, lab procedure protocols, and career advice...standard laboratory procedures. How-To Videos Screencapture walk-throughs to help guide you through Addgene's... Description Related Page Making LB Agar Plates Create plates to culture bacteria in the lab Pouring LB...with CRISPR Protocol Over-Agar Antibiotic Plating Create plates to culture bacteria in the lab Over-Agar...tips and tricks to reduce transformation time and increase efficiency Bacterial Transformation Protocol Inoculating...

...dramatically increases the chances of individual plasmids being lost from the pool and/or increasing the skewness...plasmids in a single sample. They are often used for screening, barcoding, or other high throughput multiplexed...pipettes Ice slurry (Ice bucket with ice and water to create slurry) 100 mL LB At Addgene we use premixed LB...Immediately purify them! Commercial Maxipreps rely on incremental, ordered cell lysis. E. coli cells are subject...spreading less bacteria on a given plate (example: by increasing the total number of bioassay plates used). Scaling...

...each (these can be built into PCR primers when creating a new part, with an additional 5' clamping sequence...Features: Modified RFC 10 w/ start and stop codons; Creates in-frame fusions w/ linker Thr-Gly; Use N-parts...

...Other Applications Purify Tag Visualize dCas9-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors gRNAs...another round. Figure 1: Overview of prime editing. Created with BioRender.com. The original prime editing ...

... used in the creation of the hiPS cell lines The following plasmids can be used to create cell lines with...

...used to reconstitute self-replicating microbes or recreate the diseases they cause. The standard format for...system - Wittbrodt Lab MultiSite Gateway Kit : Creating three-part fusion proteins - Planas and Santalucia...

...well in sparse/individual cultures due to lack of secreted growth factors, so not every cell type will be...cell growth, such as using culture medium with increased serum concentrations, may be optimal for different...monoclonal lines have been sufficiently expanded, screen the lines for transgene expression and/or other...phenotypes. For example, perform Western blotting to screen for lines with the highest or lowest transgene ... vectors and genome-wide libraries for CRISPR screening. Sanjana NE, Shalem O, Zhang F. Nature Methods...

...concentration of positive droplets decreases by a factor of ~2. To increase the accuracy of the titer, take...the 96-well plate sealer by gently touching the screen. Prepare the Serial Dilution Place a 48-well dilution...the plate sealer is not at temperature, touch the screen on the plate sealer to allow it to get to temperature...therefore the concentration obtained by ddPCR should decrease by a factor of 2 across the dilutions. In the ...

...Addgene has a collection of plasmids suitable for creating individual fluorescently tagged proteins to study... for FRET The following plasmids can be used to create a fluorescent fusion protein with your gene of ...

...plasmid-based technology, such that plasmids can be used to create viruses with specific genomes (Fig. 1A) . In other...associated with better memory, plasmids can be used to create viruses that encode that protein. Then, those viruses...

...Pooled CRISPR Libraries for Screening Use pooled CRISPR libraries to screen for genes involved in specific...

...the 96-well plate sealer by gently touching the screen. Preparation of the Master Mix Place a ddPCR plate...the plate sealer is not at temperature, touch the screen on the plate sealer to allow it to get to temperature... concentration of RRE positive droplets should decrease by a factor of 2 across the dilutions. RPP30 copies...below, the concentration of RRE positive droplets increases by a factor of ~2 as you progress from the higher...stays relatively even across samples (green). To increase the accuracy of the titer, calculate an average...

...at the central position. Introducing Hi-P for screening phosphorylation-dependent protein-protein interactions...NEDD4-2; 111705-8 ) is available from Addgene. Screening kinases. The mode #1 phosphosite library can also...

...temperatures, but you may need to increase your primer length and increase the Tm if you are trying to clone...Bacterial Transformation Summary PCR based cloning is incredibly versatile and allows for nearly any piece of ... 2 ligations for each plasmid you are trying to create, with varying ratios of recipient plasmid to insert...