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...to the Cas9/tracrRNA complex. D) The mature crRNA is used to guide the Cas9 complex to the target DNA which...describe the functions of the various CRISPR/Cas9 components. Please note that while this post is intended...intended to provide a general overview of CRISPR components, new Cas9 variants are being discovered all the ... thereof. This individual protein has all the components necessary to: 1. Bind to a guide RNA The guide...remainder of this post, we’ll refer to this RNA complex as the “gRNA”. The gRNA contains both the 20 nucleotide...   Now that you know about all the different components of CRISPR, It's time to start thinking about ...

...are reached, how do science labs celebrate accomplishments and their team's hard work? Do succesful scientists...each other. The Church Lab enjoys each other’s company outside of the lab by sharing meals, going on trips...! How does your lab celebrate each other's accomplishments? Any tips on making time to relax and/or enjoy... celebrating accomplishments in the lab... science labs celebrate accomplishments...

... good.  First, why are plasmids incompatible? Plasmid incompatibility occurs when multiple plasmids within...Plasmids with the same replicon are incompatible because they compete for the same replication control machinery...2012). This concept is a little more complex than simply competing for replication control factors and ...high copy plasmids, incompatibility due to partitioning is similar to incompatibility due to having the ...plasmids are incompatible? Scientists have developed a system of bacterial incompatibility groups based...belonging to the same group are incompatible. Currently, there are 27 incompatibility (Inc) groups in the Enterobacteriaceae...whether your plasmids are compatible or not.  Using plasmid incompatibility for therapeutic advantage ...

...populations and may need to be “comped” first before you apply compensation values to your current channels...do I know if I will need compensation? To know if you will need to compensate in your flow panel, you ... to apply a compensation value between the two channels (Figure 1A). Applying compensation means you are...channel and needs to be compensated out of APC-Cy7. After increasing the compensation values for APC against..., you would decrease the compensation value.   Figure 1: Compensation example with single color... be fixed with fairly low compensation values (0-50). Inputting compensation values differs between machines...point. Applying compensation to your experimental samples In order to do compensation on your experimental...

...tests. All sample comparisons Do you really need to know every pairwise comparison within your population...just perform the relevant comparisons using Dunnett’s test, a test that compares all groups to a single ...we will dive into the nitty gritty details of comparison statistical tests – when to use them, what they...your data follows. Fig. 1: Examples of comparable groups with and without equal levels of variance... discuss more in another blog. T-tests T-tests compare the mean of two groups to determine if they are...one-tailed test also works for exclusively less-than comparisons. Paired and unpaired t-tests Samples that come...occasions, there is an experimental group being compared to a standard value (not a control group). This...

...lower than that of the widely used SpCas9 and comparable to SaCas9. As seen with Cas9 orthologs, Cpf1 ...sophisticated genome-wide analysis. At many of its computationally predicted off-target sites, Cpf1 does not mediate...sites, according to Kim et al. Kim et al. also compared the ratio of total off-target to on-target modification...clinical applications require HDR over NHEJ, so comparing HDR efficiency across CRISPR nucleases and NgAgo... Up On Your Cpf1 Background Read About CRISPR Competitor NgAgo Learn How You Can Use piggyBac Vectors ...

...busy in those early years, the founders knew that company culture and enjoying your job were important parts...each other, dressing in costume, and holding competitions (ranging from cook-offs, to corn toss to trivia...in between). Eleven years later,  Addgene is a company of fifty instead of three, and we are sending hundreds...Though there still isn't time for goofing off, the company culture here at Addgene has remained paramount....Most Addgenies reply with an enthusiastic, “Our company culture! Addgene is the best place to work, we ...work, we genuinely enjoy each other’s ideas and company. The Addgene summer picnic This past weekend was... of University Park at MIT.  I’m sure at some companies, employees wouldn’t be caught dead coming into...

...choice for the job compared to T7 ligase (it was—see Potapov et al., 2018). More complex reactions benefited...tools above to complete a 52-piece assembly of a 40 kb T7 phage genome. They first compared 10-piece linear...design GGA overhang sets, particularly for high-complexity reactions? The researchers over at New England...data-driven approach might allow for an increase in the complexity. In 2018, they published a paper profiling four...2022).     Figure 1: Workflow for high complexity Golden Gate Assemblies. Image courtesy of New... three DAD tools used were key for ultra-high-complexity assembly. While the traditional rules of thumb...data-driven approach was needed to massively expand complexity. The three tools developed, outlined below, allow...

... to head NgAgo has a number of unique features compared to Cas9. These include: More targeting flexibility...sensitivity to off-targets More research is needed to compare CRISPR and NgAgo, but it seems that NgAgo has lower...

...lab. How to make your own competent cells It’s easy to make your own competent cells. If you need a high...from purchasing competent cells. However, for most everyday cloning, homemade competent cells will work...Preparation of chemically competent cells (protocol) Preparation of electrocompetent cells (protocol) At...produces 20mL of competent cells, enough for 800 transformations when using 25uL competent cells per reaction...Transformation of chemically competent cells (protocol)  Transformation of electrocompetent cells (protocol)  ...well in other competent cells) to ensure that your cells can be readily transformed. Compare the number ... time efficient. We do this by making our own competent cells and using a little-known reagent for streamlining...