... inducible Hsp68 Mammalian Melton 15508 pBS-OMP-CreFNF OMP targeting vector with Cre FNF (neo-selectable...recombination has occurred, allowing for direct comparison of Cre+ and Cre- cells. While Cre-lox recombination...Mammalian Pelczar 51267 pCAG-Co-InCreN N-terminal component of the Co-InCre system CAG Mammalian Pelczar 51268... 51268 pCAG-Co-InCreC C-terminal component of the Co-InCre system CAG Mammalian Pelczar 51273 pCAG-Roxed-Cre...Eisenhoffer 131782 pME-CreLite CreLite system components, PhyBdeltaCreC and PIF6CreN, in middle entry ...Eisenhoffer 131785 pAAV-CreLite CreLite system components, PhyBdeltaCreC and PIF6CreN, in AAV donor/transfer...Eisenhoffer 131786 pLenti-CreLite CreLite system components, PhyBdeltaCreC and PIF6CreN, in lentiviral vector...

...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...

...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 ...

... 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 ...

...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...OpenWetWare Preparing chemically competent cells (protocol)  Preparing electrocompetent cells (protocol)  Untergasser...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 ...

...approach to monitor the composition of spatiotemporally defined protein complexes." Nature communications...neighboring proteins. Thus to capture protein complexes, scientists can simply express a bait protein ...domain to allow for promiscuous biotinylation. Compared to BioID, BioID2 exhibits enhanced proximal labelling...mutants, the 35 kD TurboID, which has 15 mutations compared to BirA and the 28kD miniTurbo, which has the ...the N-terminus domain deleted and 13 mutations compared to wild type BirA. TurboID and miniTurbo both enable...interactions Split-BioID combines a protein fragment complementation (PCA) technique with proximity labelling to...vivo technique where two proteins are fused to complementary reporter proteins that can only assemble when...

... a startup company. However, like many like-minded people, I quickly realize that a company requires more... and tested in the “market” by competing in business plan competitions. After much learning, the focus...plan competition and advance investors to the next stage of diligence." In business competitions everything... goals and vision for the company; this will help avoid future complications and potential conflicts that...during graduate school and turn it into a biotech company. Transitioning an idea from academia to a startup...clearly communicate their idea at various levels of complexity and abstraction and convince people with diverse...concept development and participating in business competitions. When the team was unable to secure enough money...

...VPR fuses a tripartite complex with dCas9 to activate transcription. This complex consists of the VP64 ...sequences, CRISPR/Cas systems rely on a guide RNA complementary to the sequence of interest. Upon binding, CRISPRa...rely on fusion proteins while others re-engineer components of Cas systems themselves. SunTag, SAM, and VPR...consistently shows the highest levels of gene activation compared to other CRISPR activators, making it a popular...once), however, SAM exhibits activation levels comparable to other popular activation methods (VPR and ...initial dCas9-VP64 activator, but lower levels compared to the SAM system. Its activation levels are similar...to that of SunTag. An advantage to this method compared to other notable CRISPR activators is that it ...

...Bimolecular Fluorescence Complementation (BiFC). What is bimolecular fluorescence complementation? Bimolecular fluorescence...often easier said than done. Fortunately, a complementary method exists for visualizing protein-protein... fluorescence complementation is a method for detecting protein-protein interactions that uses split fluorescent...fragments that individually lack fluorescence. When complementary FP fragments are fused to two hypothesized protein...interaction partners (PIPs), the FP fragments form the complete FP because the PIPs they’re tethered to bring ...into close proximity (Figure 1). Formation of the complete FP results in a fluorescent signal that can be...  Applications for bimolecular fluorescence complementation BiFC has been used to directly visualize PPIs...

...known PID formats are resolvable compact identifiers, which are Compact Uniform Resource Identifiers (CURIEs... authors comply with journal policies, Addgene now provides an RRID and a resolvable compact identifier...lost (e.g. a supplier is bought out by another company and catalog number changes) the original identifier...the specific plasmids used. RRIDs and resolvable compact identifiers now exist for all Addgene plasmids!...which has the Addgene ID #87155. The resolvable compact identifier format for lenti-Cas9-VQR is the URL...to include the option of citing with resolvable compact identifier and RRID formats. Let’s go back to our...the entire line. Cite plasmid using a resolvable compact identifier and RRID “lenti-Cas9-VQR-Blast was a...

...biofilm formation. The quest for plant compounds to treat complications of diabetes Gloria Otito Izu, PhD...Her goal is to identify and isolate pure plant compounds with antidiabetic properties that can ameliorate...nephropathy, one of the major pathophysiological complications of diabetes mellitus. Recombinant human proteins...expression systems here) and the activity of the plant compounds on these proteins will be tested. Investigating...

...major histocompatibility complex, class I, A FLJ26655, HLAA HLA-B major histocompatibility complex, class...major histocompatibility complex, class II, DO beta DOB HLA-DPA1 major histocompatibility complex, class... Name Synonyms C3 complement component 3 ARMD9, ASP, CPAMD1 C5 complement component 5 CPAMD4, FLJ17816...R33729_1, SF20 C3 complement component 3 ARMD9, ASP, CPAMD1 C3AR1 complement component 3a receptor 1 AZ3B...HNFAG09 C5 complement component 5 CPAMD4, FLJ17816, FLJ17822, MGC142298 C5AR1 complement component 5a receptor...major histocompatibility complex, class II, DM beta D6S221E, RING7 HLA-DOA major histocompatibility complex...

...capture protein:DNA complexes. After capture, the DNA is released from the complex and can be sequenced...is licensed to multiple companies for sale. Before purchasing, directly compare the product information... size comparison of peptides. Independent antibody validation This method validates by comparing your ...configuration of a protein or a protein:protein complex. Due to their increased manufacturing requirements...from antibody fragments (shown in Figure 2) to completely unrelated molecules. Examples include: scFv -...nanostructures. Figure 2: An antibody (IgG, far left) compared to alternative affinity reagents (left to right...conjugating and signaling molecules, as some buffer components can interfere with the conjugate or conjugation...

...detecting multiple genomic loci, and compatible with live cell imaging. Compared to techniques like fluorescence...nuclease pair for every genomic target. Due to its comparative simplicity and adaptability, CRISPR has rapidly...approach. Engineered CRISPR systems contain two components: a guide RNA (gRNA or sgRNA) and a CRISPR-associated...Cas protein). The gRNA is a short synthetic RNA composed of a scaffold sequence necessary for Cas-binding...it meets two conditions: The sequence is unique compared to the rest of the genome. The target is present... protein and the gRNA form a ribonucleoprotein complex through interactions between the gRNA scaffold ...homology with the target DNA. Once the Cas9-gRNA complex binds a putative DNA target, the seed sequence ...

...nuclease form a complex with each individual, unique crRNA. (4) Each crRNA:tracrRNA:Cas9 complex seeks out ...a protospacer adjacent motif (PAM). The CRISPR complex binds to the foreign DNA and cleaves it to destroy...out the DNA sequence complementary to the crRNA. In Type II CRISPR systems, a potential target sequence ...after where the crRNA would bind. (5) After the complex binds, Cas9 separates the double stranded DNA target...strands near the PAM. (6) The crRNA:tracrRNA:Cas9 complex unbinds after the double-strand break. CRISPR systems...classes based on the structure of the effector complex that cleaves the genomic DNA. Type II CRISPR was...following in 2015. Class 1 (Multi-subunit effector complex) Class 2 (Single multi-domain effector) Type I ...