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...CFD scores (which range between 0 and 100) are then converted to a weight (which ranges between 0 and 1)...These values (ranging from -40 bp to 40 bp) are also converted to a weight (which ranges between 0 and...cut-to-insert distance) This gives each guide a score ranging between 0 and 1. The cutting frequency determination...specificity of a guide to the target. The CFD score ranges between 0 and 100 for each guide, with 100 being...

...O'Neill, P. S., Villette, V., Bradley, J., Wang, Z., Jiang, D., Zhang, G., … Lin, M. Z. (2024). A fast and ... J., Luebbers, A., Green, E., Philibert, C. E., Zhang, H., Layne, M. D., Wu, J. C., & Garcia-Marcos, M...diseases (Diabetes & cancer). Image adapted from Wu and Wang 2024 under a CC-BY 4.0 license. Find more neuroscience... neuroscience antibodies here! Wu, Z., Wang, W. (2024). Anti-APP [6E10-A5] in Immunohistochemistry (Human...

...prepping. Be sure to stay within the recommended range of the filter or you might inadvertently lower the... the culture medium. Improve the culture media Changing to a richer media or adding supplements can improve... low copy plasmids or slower growing strains, a change in media may be advantageous. Reduce the antibiotic...for high copy plasmids in standard growth strains range from 12-16 hours, but cultures with lower copy plasmids...oxygen for optimal growth, as insufficient gas exchange will prevent your cultures from reaching the desired...shaker speed is fast enough for sufficient gas exchange in the overnight culture. Generally, large cultures...

...players in the CRISPR field. Just a year after Feng Zhang’s lab identified Cas13a (C2c2) (Abudayyeh et al.,...end joining (NHEJ). Designing an RNA editor The Zhang lab envisioned a two-component RNA editor: a Cas13...adenosine to inosine in RNA, creating a functional A->G change. They fused ADAR deaminase domains (ADARDD) to ...introducing a C opposite the target A to be edited. This change specifies the edit to be made when multiple As ...Capabilities through RESCUE Two years later, the Zhang lab expanded the RNA editing toolkit to allow C ... their RESCUE (RNA Editing for Specific C-to-U Exchange) system (Abudayyeh et al., 2019). To avoid some...

...olds in a single evening. Grades K-5 cover a broad range of attention spans, skill levels, and STEM exposure...Chemistry activities from Quatr.us Foldscopes Steve Spangler Science The Fab Lab Middle school: supplementing... key concepts (Fig. 2). One thing I will likely change for next time is giving fewer examples and think...regulated. I did not try to sugarcoat the potential dangers of people using the technology in an unethical ...aspects of talking to 8th graders was the broad range of questions I received. I gave my presentation ...each, and each group went on its own wonderful tangents. Some groups asked about vaccines, others were...

...preferred buffer. If that’s the case, a buffer exchange can be performed using a desalting column or an...western blot. This can be due to conformational changes in the antigen or lower detection sensitivity of...antibodies may impact their quality and efficacy (Wang et. al., 2018). Aggregation can arise from a shift.... PMID: 23908655; PMCID: PMC3725456. Schwarz H, Zhang Y, Zhan C, Malm M, Field R, Turner R, Sellick C,...j.ijbiomac.2015.03.027. Epub 2015 Mar 25. PMID: 25819219. Wang W, Roberts CJ. Protein aggregation - Mechanisms,...A. Pathak, Christopher J. Roberts, Prasad S. Sarangapani, Protein aggregation, particle formation, characterization...

... dilution should fall within the range of the assay for a wide range of titers. We usually dilute our ...determine if your titers are within the expected range. We have also found that generating the standard...virus. It is important to note that the dynamic range of the ddPCR is between 1 and 100,000 genome copies...per reaction. Since AAV titers tend to be in the range of 1012 to 1013 GC/mL, you must serially dilute .../s is recommended to ensure an even temperature change among all of the droplets, but you can go as low...

...critical regions of plasmids deposited with us via Sanger sequencing, while more recently we have moved to...design, we were free to use multiple programming languages (Python, Perl, Java, R, etc.) in our pipeline.... to translate or convert programs to a uniform language.  The modern architecture of Nextflow included...Python scripts, queried our database with SQL or Django, and executed code in Jupyter notebooks using familiar...Since a Nextflow process can utilize any scripting language that can be executed on Linux, we created our ...through much trial and error to learn the scripting language, but our path forward was clear: to build one ...

...will always be that way. Things change and processes often have to change with them. If you can, set aside...concept and pare it down until you have something tangible and applicable to your situation.   Thinking Outside...creative when you don’t believe you have the power to change anything. You may not have the highest position...received, understand that people can be resistant to change. Ask your peers or your superiors to explain their...unworkable idea into a viable solution.  Anticipate Change Just because a process is working, doesn’t mean...

...increase gene expression at certain loci to create changes in phenotypes. For example, CRISPR activation has... constructs, however, Cas9 allows you to easily change the sequence targeted by the activator by simply...Voges M, Schäfer C, Spohn M, Stucka R, Hauber J, Lange UC (2016) Targeted HIV-1 Latency Reversal Using ... Habib N, Gootenberg JS, Nishimasu H, Nureki O, Zhang F (2014) Genome-scale transcriptional activation...doi.org/10.1016/j.cell.2014.09.039 Wong N, Liu W, Wang X (2015) WU-CRISPR: characteristics of functional...doi.org/10.1186/s13059-015-0784-0 Zetsche B, Volz SE, Zhang F (2015) A split-Cas9 architecture for inducible...

...molecular language shared by every living thing fit onto a single lecture slide. That shared language is the...to our Viral Vectors 101 series. Are broad host range plasmids shuttle vectors? While it’s technically... to say that shuttle vectors have a “broad host range,” these terms are typically used in different (though...(though overlapping) ways. Broad host range plasmids are those that can replicate in multiple kinds of...incredible that my cells “speak” the same molecular language as the tree outside my window! Whatever you use...

...This technique uses the trace file generated in Sanger sequencing and a simple algorithm to quantify editing... to amplify the target region of interest. Then Sanger sequence the PCR product of both an unedited population...enzyme screening is sensitive to single base pair changes, but the edit must occur at an existing restriction...simultaneously validate edits and look for off-target changes. This method requires a set of control cells to...option, you can still detect off-target effects with Sanger sequencing, provided you have some idea of where...where off target editing may occur, allowing you to Sanger sequence a manageable number of the higher probability...

...Brainbow-3.0, but with mOrange2, EGFP and mKate2 as the fluorophores; mOrange2 is the default state. In...first fluorescent protein in the construct, OFP (orange), is expressed when Cre is absent; stochastic Cre... both fluorescence and sequence overlap (coral mOrange2, jellyfish EGFP, and sea anemone mKate2.) They...proteins in the nervous system. Livet J, Weissman TA, Kang H, Draft RW, Bennis RA, Sanes JR, Lichtman JW. Nature.... Pan YA, Freundlich T, Weissman TA, Schoppik D, Wang XC, Zimmerman S, Ciruna B, Sanes JR, Luchtman JW...

... problem, how can one little change make a difference? Individual changes to our behavior can lead to ...By initiating little changes in ourselves, we can both influence others to change their behavior and we... Recycling and Innovations posts. Making small changes in your daily habits can feel ineffective or unimportant...

...Addgene saw big changes in 2022, from the launch of a new material to the redesign of a community resource...around the world, are learning to navigate the changes brought about by the pandemic. As we adjust, we...reflecting on how to help scientists in this rapidly changing environment. In particular, we are working to ...learn from, and a compelling future to plan in the changing landscape of science. We are therefore embarking...to serve the scientific community as we grow and change to meet its ever-evolving needs. For now, we have...

...., Liu, X., Cai, Y., Kou, J., Wang, J., Li, L., Peng, S., Xu, Z., Zhang, W., Qiu, Y., Wu, Y., Ye, C., ... material, and variations between serotypes can change how the viral particle sneaks its way into cells...Tabar, M. S., Metierre, C., Parsania, C., Song, X., Wang, G., Su, X.-D., Bailey, C. G., & Rasko, J. E. J....://doi.org/10.1038/nature16465 Tervo, D. G. R., Hwang, B.-Y., Viswanathan, S., Gaj, T., Lavzin, M., Ritola...Lindo, S., Michael, S., Kuleshova, E., Ojala, D., Huang, C.-C., Gerfen, C. R., Schiller, J., Dudman, J. ...

...Proteins For Molecular Biology Experiments by Jennifer Tsang Chromoproteins are a subset of the fluorescent ...many molecular biology methods such as allelic exchange, FRET, and synthetic biology. 7. 25 Work From...From Home Tips for Scientists by Jennifer Tsang These tips were written early on in the pandemic. Later...Quantification Methods to Consider by Jennifer Tsang The nanodrop vs. spec debate dominates DNA quantification... Jennifer Tsang...

...expression vector for CRISPR nuclease, SpCas9 from the Zhang Lab. It's name, though a little confusing to the... is a modified version of CAG promoter that the Zhang lab has tested in a variety of mammalian cell lines...component of most CRISPR systems and generates double strange breaks at target sites 5' of the 5'NGG3' PAM sequence... can be modified for viral trasduction (see the Zhang Lab's Nature Protocols Paper for more information... all of these variants, as well as links to the Zhang Lab's Nature Protocols Publication describing them...

...domain is interesting because by changing certain amino acids you can change the specificity,” explained Zhanar...she hopes will change the intracellular localization of RNA. “You can think of changing bases to other...binds a consecutive base in its RNA target. By changing amino acids, it’s possible to produce a protein...

...graduate student Silvana Konermann first entered Feng Zhang’s lab at MIT, the use of customizable DNA-binding...new. Now, Konermann and her colleagues including Zhang and Mark Brigham have taken the technology to another...literally at the flip of a (blue) light switch. Zhang’s team devised the plasmids now available in Addgene...introducing the optogenetic tool, Konermann and Zhang showed they could package LITEs in viral vectors...specific epigenetic modifications in the genome,” Zhang said in MIT’s press release. In an accompanying ...