...multi-protein complex, stabilized promoters can eliminate variability between the different components. When ... origin of replication will be overrepresented compared to those further away. As a result, variable gene...located near the origin will be over-represented compared to those further away. While constitutive promoters...gene expression during shifts in growth rate as compared to a constitutive promoter. To simulate the increase...

...available in rAAVs!   Figure 1: Diagram of AAV components      Cargo capacity Empty rAAV has a cargo...conventional single-stranded AAV (ssAAV) and a self-complementary AAV (scAAV) vector. Full-length ssAAV vector...2.5Kb) and you need expression in a hurry, self-complementary AAV (scAAV) may be an option (McCarty et al....take the next step – selecting your AAV and AAV components (serotype and promoter) for use in your experiment... E. Monahan, and Richard J. Samulski. "Self-complementary recombinant adeno-associated virus (scAAV) vectors...

...below) Because off-target sequences have less complementarity to the gRNA, they have a higher propensity ...SpCas9-HF1 generated fewer off-target cuts when compared to WT SpCas9 across a variety of genomic sites...M694A/Q695A/H698A). This new variant displays comparable on-target activity and similar or decreased off-target...off-target activity across the sites tested when compared to eSpCas9(1.1) and SpCas9-HF1. The HypaCas9 plasmids... Hiroshi, et al. "Crystal structure of Cas9 in complex with guide RNA and target DNA." Cell 156.5 (2014...

... The destination vector contains sites with complementary overhangs that direct assembly of the final ...recognition site and permits DNA fragments with compatible overhangs to be efficiently assembled. Scientists...inverse orientation. This allows for multiple DNA components (promoters, genes, terminators, etc) to be assembled... reaction. For detailed Golden Gate protocols, complete with helpful tips and tricks, see The Sainsbury...system is an incredibly powerful tool for cloning complicated constructs in a single, high-efficiency step....

...audience’s eye and draw people in. That can be accomplished with an exciting title and attractive figures...in less than a couple of minutes. How does one accomplish all of that? What follows are some tips to help...logical progression of your work, and you can accomplish this by employing a neat and organized format...length, and will be more difficult to read. The Thompson Rivers University Library provides useful tips...want to make sure that the color of your font is compatible. Use high-contrast between these colors so that...

... don't do anything. Viruses are complicated and they perform complex biological tasks, both of which ...undergo DNA rearrangements. Depending on the composition of the viral vector and the repeat sequences ...perform a diagnostic restriction enzyme digest and compare it to some reliable controls. 2. Viral stocks may...ensuring viral production is to perform PCR against components of the viral vector using the harvested viral...

...benefits and drawbacks of different methods (for comprehensive protocols for all of the methods discussed here...quantified using qPCR primers targeting specific viral components such as LTRs, gag, WPRE, antibiotic resistance-genes...genomic DNA isolated, and primers targetting viral components or the transgene itself are used to quantify ...critical that standard conditions be used when comparing different batches of virus. In addition, some ...PMID: 19300443. 2. Geraerts, Martine, et al. "Comparison of lentiviral vector titration methods."BMC biotechnology...

...about a month and a half, without the need for complicated cloning of homology arms. MMEJ: An Introduction...HDR and NHEJ, please see the linked blog posts. Compared to its counterparts NHEJ and HDR, MMEJ doesn’t...strategy looks very similar to HDR, but it is accomplished using much smaller regions of homology, which...efficiency of PITCh-based genomic insertion would compare to CRISPR sticky-end insertion using the nuclease...Genome-Editing Based Approach for Mapping Multiprotein Complexes in Human Cells.” Cell Rep. 13: 621-633. PMID: ...

...Revolution Bioengineering.  Last year we started a company. Revolution Bioengineering is two responsible adults...attend conferences makes it very challenging to compete in today’s tough academic or industry job market... this technology and angry at the agriculture companies that produce GMO food plants. When a press piece... – although we didn’t figure out how we would accomplish the color change until a few months later. RevBio... of our business, because we are building this company, and that’s what we want it to do. It’s exhilarating...

... in other lanes contaminating your sample. To accomplish this, it is best to skip lanes between samples...

...trace file (.ab1 file) provided by your sequencing company or core facility The corresponding primer sequence...

...reagents is important to ensure the accurate composition of your solutions and buffers. Now that you have...

...plasmids, they are generally electroporated into electrocompetent cells to ensure the highest possible transformation...use for your transformation(s). The amount of competent cells you need total and per transformation may...liquid culture, all of the bacteria can grow and compete in the same container, making it easier for a smaller...be its own colony, and will not be in direct competition with other colonies containing smaller, recombined...used for libraries that are similar in size and composition as examples. Table 1 provides an overview of ...

...expression baseline. Copy number is then determined by comparing results to a standard curve. For CNV assays, it...gives an absolute value of DNA concentration. By comparing the number of positive droplets for the gene of... through concentration values, rather than by comparison to a standard curve, and most results can be ...determination to move forward with your experiment?  Complexity of the gene of interest Some gene sequences ... S., Hollox, E. J., & Boon Peng, H. (2015). A comparison of assays for accurate copy number measurement...

...you’ll want to ensure that your buffer components are compatible with your conjugate and conjugation chemistry...the assay. Where possible, it can be useful to compare recombinant antibodies to their parental hybridomas...majority of antibodies are still equally effective compared to those aliquots frozen immediately post-harvest...Chen E, Haidar JN, Srivastava A, Goldstein J. Comparison of predicted extinction coefficients of monoclonal...

... takes 1-1.5 hours to complete, while this syringe-based method can be completed in 20 to 30 minutes. ...PCR buffer, where DNA or RNA can be amplified. Compared to a commercial magnetic bead-based kit, this ...this did not reduce cloning efficiencies.  When compared to fresh columns, regenerated columns had similar... reduce the number of spin columns you need to complete your research. Do you know of other nucleic acid...

...feel comfortable speaking on topics that would compliment the STEM curriculum. As it happened, I’d met ...simplistic and I would insult the students. Too complicated and I would quickly lose their interest. I won...careful to keep it relatively simple. I honed the complexity of the talk by running it by the teachers and... For CRISPR, I focused on the fact that, like computers and the internet before it, genome engineering...posts Resources on Addgene.org Check out our comprehensive CRISPR guide Find CRISPR plasmids and resources...

...one such technique and has been used to produce complex fluorescent reporter constructs. These fluorescent...gene-expression sequences, and more. By combining these components, one can generate constructs to visualize cell...terminus or the 3’ terminus of the ligation product. Comparison to other high throughput cloning strategies There...modules, because neighboring fragments require compatible cohesive ends Engler et al., 2008 Gibson Cloning...facilitate the construction of plasmids with many components. The creators of the MXS-chaining method have...

...will link biomolecules to the gel and add gel components to polymerize. Step 3: digest the sample to allow...labels you could use. But many ExM protocols are compatible with conventional antibodies, chemical dyes, ...and least flexible one. Tissue expansion is not compatible with in vivo applications and there’s just not...isotropic expansion depends on how well the gel components penetrate the sample, how well the gel polymerizes... valuable tool, especially when paired with complementary experimental approaches. Since it arrived on...

...collaborations, and commitments to and proof of compliance with the grantor’s requirements as well as state...reports are submitted and project evaluations are completed. To enter and advance the profession, you can ...Professional Development Strategy to increase your competencies and skills in the grants profession. Your professional... and the candidate’s knowledge of the 8 core competencies through a multiple choice exam.  The bottom ...grant professionals and to build your skills and competencies. Becoming a grant professional is rewarding ...