... PMCID: PMC5312033. 2. Dewari PS, Southgate B, Mccarten K, Monogarov G, O'Duibhir E, Quinn N, Tyrer A,...

...: Immunofluorescence staining for CD31 (green), CARD14 (red), and pNFkB (blue) in lesional skin from a...

...fluorophores and their emission spectra. Some fluorophores carry their emission peaks within their names, for example...

...strategies are presented (Fig. 1) which can be carried out using Addgene plasmids. His-tag labeling reagents...

...Overlay Select Science Career Guide November 2018 | First Edition About Science Career Guide November 2018...About The guide includes information on the many career options in science, advice on mentoring and management...

...plasmids from this paper at Addgene. 12. Ronda, Carlotta, et al. (2016). "CRMAGE: CRISPR Optimized MAGE...

...Restriction enzyme cloning also leaves behind a short scar in the DNA sequence and can be time consuming compared...entire cloning step (digestion and ligation) can be carried out in one reaction with a single type IIS restriction... the final product, with no undesired sequence ("scar") retained. A disadvantage of type IIS assembly ...

...stored at 4 °C for up to 2 months. After 2 months, discard the tube and thaw a new working stock. The optimal...the transfection mix onto the 10 cm plate, being careful not to disturb the cells. Incubate the cells for...the following morning. The following morning, carefully aspirate the media. Replace the media with 10 ...

...editing is precise and versatile, but it requires careful optimization to achieve the best efficiency. The...

...polymerase and a blend of dTTP/dUTP to minimize carryover contamination. The master mix should also contain...DNase I to eliminate any contaminating plasmid DNA carried over from the production process (DNase does not... 3 Pro-Tip To help stabilize the standards add carrier DNA to a final concentration of 4 ug/mL to each...

...This blog post was originally written by Caroline LaManna, published Mar 8, 2016. The updated and expanded...

...agar. This usually takes 1-2 minutes. Critical Be careful not to rip or shred the agar. Do so by gentle spreading...Pushing motion is better than pulling motion Take care not to split or gouge agar during the scraping process...library even in the face of a recombined fraction but care must be taken to perform adequate NGS based QC to...

...spills produced during the protocol are cleaned up. Discard the weighing boat or weighing paper in the trash...weighed is not biohazardous. If it is biohazardous, discard the weighing boat or paper in the biohazard solid...

...Biolabs: Introduction to AAV University of South Carolina's Viral Vector Core AAV Overview Publications Viral...efficient transduction independently of DNA synthesis. McCarty DM, Monahan PE, Samulski RJ. Gene Ther. 2001 Aug...

...that can be used to titer lentiviral preparations carrying a fluorescence marker....can be used to titer lentiviral preparations that carry a fluorescence marker. Determining the titer of ...

...even those designed for mammalian cell expression) carry both a bacterial origin of replication and an antibiotic...overnight. Tips and FAQ How can I save time when carrying out transformations? If you are not concerned ...

...sure to check whether reagents are available to carry out your experiment in your particular model organism...save your lab valuable time and resources when carrying out CRISPR experiments. Read more about how to...

...touch the sides of the container with the pipette. Discard the pipette tip by holding the pipette tip over...If pipetting biohazardous materials, be sure to discard into a biohazardous waste container. Conclusion...

... process is shown graphically in the following cartoon, in which we are adding EcoRI and NotI sites to...Verify your Plasmid by Sequencing PCR based cloning carries a much higher risk for mutation than restriction...

...microcentrifuge tube. Centrifuge for 5 min at 100 x g . Carefully remove supernatant. Resuspend cell pellet in appropriate...transfer stack. Set the Top Stack to one side and discard the white separator. Keep the Bottom Stack in the...

...results - for a given plasmid, think of each cell carrying it as being a biological replicate for that plasmid...

...restriction sites from the final product (no "cloning scars"). A “stuffer” sequence allows for electrophoretic...

... When you are finished spinning your samples, carefully remove them from the centrifuge so as to not dislodge...

...single reaction Ligation Independent Cloning (LIC) Scarless cloning with Type II restriction enzymes and T4...

...almost any two fragments regardless of sequence. No scar between joined fragments. Fewer steps. One tube ...

...the stain. Wash cells 3x with 1 mL of PBS, being careful not to disturb the colonies. Count the colonies...

...unsure of the correct orientation of the gel. Carefully remove the comb from the gel. Rinse each well ...

...contain a monoclonal population and should be discarded. Pro-Tip When scanning the plate, be sure to look...