...improve solubility and stability (Figure 2).      Figure 2: Crystal structures of CRBNmidi alone...fluorescent tags in Caenorhabditis elegans. Genetics, 228(2), iyae126. https://doi.org/10.1093/genetics/iyae126...are essential tools for visualizing biological processes in vivo. However, mScarlet’s slow maturation rate...like the C. elegans embryo, where developmental processes happen within a few hours. In order to use the...addition to the toolkit for visualizing cellular processes in C. elegans: mScarlet3 for applications needing...

...presented in a series of quick (10 minute) or very quick (2 minute) talks so we got to hear from everyone. Addgene...evolutionary tree at least 6-7 times and on at least 2 separate continents. Mimulus (Andrea Sweigart) Monkeyflowers...Organisms, Part 1 Five Popular Model Organisms, Part 2 ...’ll choose to study Crepidula atrasolea or Prochlorococcus with your next project. Gymnotiformes and Mormyridae...evolution in immunology, neurobiology, and more. Prochlorococcus (Penny Chisholm) This cyanobacterium is the...need to grow to get a recombinant because this process is expensive. They are using a cool fluorescent...

... Figure 2: Ironing boards. Good as a WFH desk, and as a video control center.   Cycle 2: Zooming ...ironing board, giving it the perfect height (Fig. 2). We recorded video using a simple, external video...doesn’t, and trying all over again. Here’s that process!   Figure 1: The Design-Build-Test-Learn...gear, and walking everyone through the recording process was going to be time consuming and difficult. Finding...in the future, but we were finally refining the process down to a place I could be happy with it. Cycle...

... environmental sustainability in mind (Table 2). Table 2: Sustainability features highlighted in plasmid...debris”; and then clean-up the plasmid DNA (Figure 2). For most kits, the steps for lysing cells and removing...together — no mixing and matching!   Figure 2: Plasmid isolation workflows are similar across kits...that can arise during the plasmid purification process. One is the possibility of damaging the plasmid...carrying over contaminants from the plasmid prep process into the final plasmid eluate. Possible contaminants...contaminants can sneak in from almost any step of the process and include things like genomic DNA, RNA, proteins...

...delicate axonal and dendritic processes not previously visible with Brainbow-1 and -2. The general structure ...Brainbow mouse neural labeling system. Brainbow-1 and -2 have been further refined to create Brainbow 3.2. ...M-CFP expression; these deletions are defined using 2 loxP variants (only identical loxP sites can mediate... each fluorophore. Third, Brainbow-1 and Brainbow-2 each contained a “default” state; for example, Brainbow...

...indicating primer dimers. (c) A double peak indicating 2 amplicons. Image credits (respectively): Zuzanna K...details for reproducibility’s sake!   Fig. 2: Setting up triplicates in multiple configurations...used method is the Standard Curve method. See Table 2, (adapted from Wong, 2018) for a comparison of different...Statistically defined log-linear phase   Table 2: A comparison of different qPCR analysis methods. ... for RNA quantitation. Biotechniques. 2000 Aug;29(2):332-7. doi: 10.2144/00292rv02. PMID: 10948434.Taylor...Getting your RQ value isn’t the end of your analysis process. You’ll still need to do statistical analysis to...Gene Expression Analysis During iPS Reprogramming Process. Sci Rep 8, 8716 (2018). https://doi.org/10.1038...

...AAVs in the clinic Thus far, AAV vectors AAV1, AAV-2, AAV1-AAV2 hybrids, AAV-6, AAV-7, AAV-8, AAV-9 and...neutralizing antibodies against some AAV types, AAV1, 2, 3 and 5 are thought to be as high as 70% among the... 10.24 (2003): 1999-2004. PubMed PMID: 14566358.  2. Hirsch, M. L., et al. "Self-complementary AAV mediates...vectors Learn about Addgene AAV Quality Control Process Resources on Addgene.org Find ready-to-use AAV...

... is the genetic test used for sickle cell anemia (2). Southern blotting, in conjunction with restriction...program) can then reproduce the DNA sequence (Figure 2). Accuracy and accessibility led to the widespread...biology98.3 (1975): 503-517. PubMed PMID: 1195397. 2. Chang, Judy C., and Yuet Wai Kan. "A sensitive new...recording DNA abundance throughout the amplification process, rather than just at the end as in standard PCR...became the workhorses of molecular biology." Proceedings of the National Academy of Sciences 102.17 (2005...restriction endonuclease of Hemophilus influenzae." Proceedings of the National Academy of Sciences 68.12 (1971...sequencing with chain-terminating inhibitors." Proceedings of the National Academy of Sciences 74.12 (1977...

...China, Korea, Japan, Brazil, India, and Mexico. Table 2: Addgene distributors Distributor Partnership...distribution after we partnered with them (Figure 2). In others, the distributor helped solve an issue...temperature for the duration of their journey. Figure 2: Annual distribution of items to countries with a ...import and export regulations, differing customs processes, and the distances the packages needed to travel...addgene.org. Currently, we’re evaluating our depositor process to identify and reduce barriers for scientists ...

... than control, 3; same as control (Cetn1/sgRNA1), 2; darker than control, 1; very dark). Brighter fluorescence...you can hold off on genotyping until the pups are 2-4 weeks old and use the ear tag method (creating different...PMID: 24253446. PubMed Central PMCID: PMC3875854. 2. Mashiko, Daisuke, et al. "Generation of mutant mice...a post that goes into the mouse genome editing process in a bit more detail in the coming weeks, but, ...Target Mouse Genes To begin the in vitro screening process, you should first develop primers to the approximately...

...phages but to also destroy the phage upon re-exposure (2, 3). More specifically, specialized Cas proteins snip..., accessed 25 November 2013. Author: James Atmos (2). Many CRISPR/Cas moieties carry out these functions... also disrupted to elucidate their role in the process. The results of these studies led scientists to...sequence. Separate Cas proteins then express and process the CRISPR loci to generate RNA, which guide a ...

...identifying the resources needed to accomplish that goal. 2. Identify the tools in that timeline Avoid writing...based on experimental timelines and specific aims. 2. Then make decisions based on financial values. All...Would I be able to get access to these materials?” 2. Always negotiate for a win-win decision An institute...value that helps you determine whether or not to proceed with a used or new item. To further illustrate ...we mentioned previously.  There is value in the process of creating a budget; you can share your budget...

...Cpf1 is a single RNA-guided endonuclease of a class 2 CRISPR-Cas system." Cell (2015). PubMed PMID: 26422227... Find plasmids from this publication at Addgene. 2. Kleinstiver, Benjamin P., et al. "Genome-wide specificities...Cpf1 Is a Single RNA-Guided Endonuclease of a Class 2 CRISPR-Cas System. Cell. 2015 Sep 23. pii: S0092-8674...overhangs. Exploring the efficiency of these two processes will help us better understand where Cpf1 fits...

...with TALENs, in collaboration with the Porteus lab [2]. With the advent of CRISPR/Cas9, we began to explore...nucleases. Genome Res. 2010 Jan;20(1):81-89. PubMed. 2. Bauer DE, Kamran SC, Lessard S, Xu J, Fujiwara Y,...following video to learn Daniel and Matthew's procedures. The video details how to transfect CRISPR plasmids...

.... Let's look at an experimental example in Figure 2. Figure 2A shows the sample prior to compensation.... line with the other populations.   Figure 2: Compensation example based on experimental sample..., you will see the populations move, as in Figure 2, following compensation. Figure 3: Flow plot...In flow cytometry, compensation is the process of correcting spillover from one fluorescent channel to...

...Marcotte EM, Voigt CA. Nature. 2005 Nov 24;438(7067):441-2. PubMed. Find the plasmids from this publication ...Levskaya A, Voigt CA. J Mol Biol. 2011 Jan 14;405(2):315-24. doi: 10.1016/j.jmb.2010.10.038. Epub 2010...fact, precise light-based control of biological processes is highly useful in other fields, including synthetic...to directly activate and inactivate biological processes. Chemical effectors are suboptimal for a number...intensity and wavelength can be used to study dynamic processes. Conversely, varied input patterns may also be...

...design primers to detect the presence of your insert; 2) set up a standard PCR reaction (primers, dNTPs, polymerase...strategies for primer design: 1) insert-specific primers, 2) backbone-specific primers, and 3) orientation-specific...negative clone (-) will not. Backbone-specific primers (2) give larger sized products for positive clones (+...sequencing to verify the insert sequence before proceeding with your experiment. Shorter amplicons tend...

...the system outlined in Schmid-Burgk, J.L., et al (2). In this post, I’ll cover two great applications ...molecularly evolved from mNeptune2 through 33 mutations and 2 deletions. CyOFP1 has a higher quantum yield, brightness...luciferin 24 hours later demonstrated a more than 2-fold brighter signal from Antares as compared to Fluc...PMID: 22894855. PubMed Central PMCID: PMC3501149. 2. Schmid, J.L., et al. (2016) CRISPaint allows modular...Here, NLuc internally excites the fluor through a process known as bioluminescent resonance energy transfer...

... intensity of the cell’s illumination (see Figure 2). H describes the maximum signal strength, while W...SSC-A, so most people use that setting. Figure 2: As the cell passes through the laser beam and causes...scale in proportion (while H and W do not; see Figure 2), single cell events will show up in a straight line...heterogeneous cell population, for example from a processed piece of tissue, where cells are present that ...structural complexity. After several steps of tissue processing prior to running my flow analysis, I can however...interest). Let’s double-click inside our gate and proceed to the next plot! Pro tip! If you only get very...

...resistance to the herbicide chlorsulfuron (Homo line 2) or wild type (WT), two weeks after herbicide was ... CLIPf tag and a C-terminal SNAPf tag; see Figure 2), the authors developed a cost-effective expression... when associated with microtubules. Figure 2: CLIP-SNAP fusion proteins for single-molecule FRET... inserted. B) Protein expression and labeling procedure. Adapted from Smith et al. 2024 under a CC-BY ...