We narrowed to 718 results for: des.1

...learn more about each reporter or check out table 1 for a summary. HRP HRP tagged RANbodies don’t require...with a standard primary/secondary antibody. Table 1: Summary of the Key Features of HRP, Spaghetti Monster...NIH’s Developmental Studies Hybridoma Bank (DSHB). 1 ml of antibody-containing hybridoma supernatant is...overcomes this limitation and allows for the flexible design and small scale production of antibodies. Problems...Their His tags allow for further purification, if desired, but it's not required for their use. Each RANbody...

...current state (Green when off, dark when on). Figure 1: Photoswitchable MEK1 (psMEK1). Is it possible to ...Below are a few examples demonstrated in Zhou et al. 1. Testing kinase inhibitors Zhou et al examined the...the protein and pdDronpa are preserved. References 1. Zhou, Xin X., Linlin Z. Fan, Pengpeng Li, Kang Shen...Solution: In Zhou et al, the Lin lab present a generic design for creating photoswitchable kinases and use MEK1...create other photoswitchable kinases? The general design of psMEK1 also worked for creating three other ...

... products). I picked this post for a few reasons: 1) I think phage are super cool and have considered ...Connect the Research Community Grad School Advice Part 1: Picking a Lab and a Project That’s it for now. As...record breaking month and to find other posts that deserve a second look. 5 Great Apps for Lab Life This very...Stacey from Eastern Tennessee State University and provides a brief rundown of Sean’s top five smartphone ...Jessica Sacher from the University of Alberta and describes a technique you can use to find phage proteins...molecular biology tool, P1 phage. 2) The post describes an ingenious but simple technique to screen for...that, while affecting a small number of people, deserves our attention and has huge potential for the development...

...cleavage of the transcript.   Figure 1: A predicted conserved secondary structure and sequence conserved...prokaryotic terminators and deposited several with Addgene (1). Prokaryotic polyadenylation Although mostly thought...constitutive RNA transport element lead to enhanced HIV-1 Gag expression in a context-dependent manner. Nucleic...Plasmids designed to express genes in a given host cell type are generally broken down into two broad...post-transcriptional additional of multiple adenine (A) nucleotides to the tail of a messenger RNA transcript. The...structure of the hairpin-DNA complex is thought to destabilize the transcription complex, initiating cleavage...transcription of the template and initiating the desired cleavage event, although some engineered terminators...

..., check out Addgene's CRISPR Guide. Figure 1: Like this jar of jellybeans, a pooled CRISPR library... selecting a CRISPR library for your experiments. 1.  What species are your cells derived from? Currently...a different gene. Libraries may be available in a 1-plasmid system, in which Cas9 is included on the gRNA-containing.... Individual gRNAs are designed in silico using publicly available gRNA design software and synthesized... be independently validated to ensure that the desired modification produces the phenotype you screened...in the first place. With the proper experimental design and validation practices, CRISPR libraries can ...CRISPR Plasmids Read our CRISPR Guide Find tools to design your gRNA ...

... the samples stored in our lentivirus tubes (Fig. 1). However, once thawed, there was a substantial reduction...in the pH of samples stored in our AAV tubes (Fig. 1). It’s important to note that the change in pH only...using the thawed virus within 24 hours. References 1. Murphy BM, Swarts S, Mueller BM, van der Geer P, ...shipment processes. After reading a publication describing how short term storage on dry ice can shift sample...

...s pLKO.1 protocol. Pro-tip: this plasmid grows more slowly than standard plasmids. Find pLKO.1 - TRC cloning...experiments.  Find the scrambled shRNA vector. pLKO.1 - TRC cloning vector - This vector from the David ...2A-Puro (PX459) - This mammalian expression vector encodes both SpCas9 and your gRNA. It’s the former version...

...step to colony PCR is designing primers. There are 3 strategies for primer design: 1) insert-specific primers... less money. The key steps to colony PCR are: 1) design primers to detect the presence of your insert;...the three primers previously described. When using insert-specific primers (1), positive clones (+) will... second option is to design backbone-specific primers. These primers are designed to anneal to sites that...things to consider when performing colony PCR.   Designing colony PCR primers The first and perhaps most ...-specific primers: Insert-specific primers are designed to anneal to an insert-specific sequence. This...backbone but that contain different inserts. When you design primers to anneal outside the cloning site, it ...

...active for greater than 50 generations. Figure 1: Mobile-CRISPRi's modular components. Image from Peters...on a non-selective plate. Most bacteria tested had 1 transfer in every ~100 to 1,000 colonies, but some...including E. faecalis and P. mirabilis, had only ~1 transfer per 1,000,000 colonies. Higher transfer rates... human gut flora. Guides targeted essential and non-essential genes with two guides per gene. The library...expressing guides that target essential genes were less represented while bacteria containing guides targeting... Jason Peters, Oren Rosenberg, and Carol Gross provides a streamlined suite of tools for using CRISPRi...existing bacterial CRISPRi methods. First, its modular design allows for customization with strain-specific promoters...

...with a membrane-impermeable biotin substrate (Figure 1). Biotinylated proteins were isolated, identified ...transporters, ion channels, GPCRs, and more.  Figure 1: Workflow for cell-surface protein labeling and identification...564–571. https://doi.org/10.1038/s41586-022-05682-1 Zhang, Y., et al. (2025). Probing condensate microenvironments... and synthetic biology. Nature Communications, 15(1), 6563. https://doi.org/10.1038/s41467-024-50767-2...required, through mitochondrial transfer. In the two decades since this transfer was first proposed, researchers..., 2025). These new sensors use quite different designs, leading to unique advantages. While most red GECIs...two-photon microscopy, and fiber photometry. Plus, both designs offer promising templates for future improvements...

...unlimited supply of identical antibodies (See Fig 1.). rAbs are a relatively new technology that takes...conveniently modified and engineered.   Fig. 1: Antibody production methods    NeuroMab and others... the signal. If the exposure time is long (e.g. > 1/7s) then the signal is weak and vice versa for a strong...perform differently over time), compatible with the desired applications (such as immunohistochemistry (IHC...antibodies, we have developed a criterion that includes accuracy, strength, and reproducibility. We test...

...expected in another. You’ll need to validate that points 1 and 2 hold true under your specific experimental conditions...application and available resources.     Figure 1: Table summarizing the IWGAV five proposed antibody...like immunohistochemistry on human tissues, when (1) other strategies may not be feasible and (2) large-scale...validation: A view from the mountains. New Biotechnol 45:1–8. https://doi.org/10.1016/j.nbt.2018.08.002  Uhlen...to be present and recognizable by your antibody despite the manipulation. Additionally, not all sample ...large deletion that removes the entire exon that codes for the epitope recognized by your antibody. You...specific, then you expect that the bulk of the peptides identified to be from the target protein. This...

...homologies. Abbreviated protocol for PITCh Step 1: Generate microhomologies in the PITCh vector ~20...editing possibilities MMEJ can enable. References 1. Sakuma, Tatsushi, et al. (2016).  “MMEJ-assisted ...CRISPR-Cas9 with the PITCh systems.” Nat Protoc. 11(1): 118-33. PMID: 26678082. Find the plasmids from ...such a system is likely to introduce additional nucleotides flanking the insertion. If the DNA ends anneal...MMEJ may also introduce, substitute, or delete nucleotides in addition to the expected deletion, but the...Building on the lab’s previous work, Sakuma et al. describe a detailed protocol for MMEJ-mediated knock-in...PITCh vector via In-Fusion or SLIC cloning. Step 2: Design an insertion locus-specific gRNA The gRNA should...

...process is explained more fully in the animated Fig. 1 below. Normalization is a fundamental and often understated...plexWell™ libraries is well suited to large-scale de novo assemblies of short constructs like plasmids...all over the world. The rest of this piece will describe plexWell™ in more detail, and how we are using... containing fragments carrying sample-specific barcodes. Rather than continuing to process 100s to 1000s...

...cell with the images from the Allen Cell Explorer (1). When studying the function of a protein or its role...various subcellular structures. Please see Figure 1 for commonly used markers for major mammalian organelles...Subcellular Localization Poster here! References 1. Allen Institute Plasmid Pages 2. Cooper GM. 2000....interest from its production site to its final destination. As an example, most secreted proteins are produced...pathway to their subcellular or extracellular destination. At the end of their life cycle, proteins may... about the subcellular structures your protein resides in and potentially which other proteins it forms...in Figure 2 (adapted from 4) - Dunn et al. 2011 provides an in-depth review of quantitative colocalization...

...glassware should be treated with a DEPC solution for 1 hour, and autoclaved to remove residual DEPC. Alternatively...more large, expensive ultracentrifuge.  Figure 1: An outline of the different steps in RNA extraction...extraction: twenty-something years on. Nature Protocols 1:581–585. https://doi.org/10.1038/nprot.2006.83 Additional...Allewell and Sarma, 1974). Additionally, ThermoFisher provides a protocol on how to integrate RNAlater® with ...

...Listen to the interview with Darrell Kotton Chapter 1: Introduction to Darrell Kotton and the Boston University...Unit, the Pulmonary Consultation Service, the Alpha-1 Center, and the Pulmonary outpatient clinic, all at...University Center for Regenerative Medicine. Darrell describes his career goals and what drives him as a praticing...

...some technical obstacles when working with virus: 1. Viral vectors undergo DNA rearrangements. Depending...approximately 20–50% of the packaging cells after 1–3 days of selection. A lower percentage of surviving...up-to-date tips for your virus experiments. References 1. Anthony, Simon J., et al. "A strategy to estimate... try amplifying the viral vector with bacteria designed to minimize rearrangements such as NEB Stable ...production of virus. Alternatively, if the virus encodes a fluorescent protein, the packaging cells could...working with, there’s a lot that goes into virus design, production, and transduction. Once you’ve harvested...

...tethered to bring them into close proximity (Figure 1). Formation of the complete FP results in a fluorescent...detected by fluorescence microscopy. Figure 1: How BiFC works. A split FP (e.g., yellow fluorescent... protein interactions in living cells. Nat Protoc 1:1278–1286. https://doi.org/10.1038/nprot.2006.201 ...This post was contributed by Patrick Miller-Rhodes from the University of Rochester Medical Center.  ...plasmids for your experiment at Addgene! How to Design a BiFC Experiment Much like FRET, BiFC experiments...quantification (Kerppola, 2006, 2013). These are described in more detail below: Fusion protein construction...each PIP and empirically test which combination provides the best fluorescent signal. As for linker sequences...

... high brightness and fast maturation time (Figure 1). See the paper for more details on small differences...mGold2t depending on the application. Figure 1: Super-resolution imaging of actin filaments with ...proteins for extended imaging. Nature Communications, 16(1), 3241. https://doi.org/10.1038/s41467-025-58223-5...., 2025). By co-delivering TRV1 with TRV2 that encodes ISYmu1 and an ωRNA under the same promoter, they...different applications require careful selection or design to ensure reliable readout and avoid biases. The...ex vivo HSC expansion cultures. A) Experimental design. B) UMAP of cell states in ex vivo expansion cultures...only at excitatory synapses; however, its modular design allows components to be swapped out, potentially...

...environments such as live cells and tissues (Figure 1). These flexible strategies give you the best of both...majority of in-cell labeling strategies have two parts: (1) a genetically encoded “tag” expressed as a fusion...Follow him on twitter @rhodamine110.   References 1. Liu, Zhe, Luke D. Lavis, and Eric Betzig. "Imaging...individual molecules. Poor photon budgets are a widespread issue with fluorescent proteins—even the thriftiest...student searching couch cushions for spare change, desperate to extract a few more photons from a sample. Chemical... the complexity of the system. A particularly desirable property for small-molecule labeling strategies...size, brightness, and cell permeability. In past decades, further refinements to this established dye structure...

...a rate of up to 1 per 500-1000 bp. In comparison, the DNA replication error rate is 1 per 109 bp. Unlike...common 30bp overlap sequences (Sites #1-5). Each overlap site encodes a unique restriction enzyme site and...Transcriptional Output-TANGO (PRESTO-TANGO) to screen the NCC-1 library of approved drugs against the entire kit; ...First, GMAP-compatible backbones for lentivirus LV 1-5 and retrovirus RV 2-5 were constructed. The authors...is multiplex automated genome engineering (MAGE) (1). MAGE uses recombineering (2) to simultaneously incorporate...of GFP (SEP) by a 9 amino-acid linker (see Figure 1). The key to the biosensor lies in pH: DsRed is relatively...inactivate SEP and leave only DsRed to fluoresce (1). With variants that can be targeted to specific cellular...

...-public settings. Ask over coffee.  Have regular 1:1 meetings with team members (don’t become known for...communication between all members of a team. As described here in a 2012 Intuit blog post, a study coming...

...Recently I learned that Addgene’s pLKO.1 cloning protocol is viewed around 3,000 times each month. I ...on the Addgene website in December 2006, the pLKO.1-TRC cloning vector (deposited by David Root of the...would the thousands of other scientists using pLKO.1 variants learn about those updates? This is a problem...the site. Like Labguru, Quartzy offers software designed with scientists workflow in mind and can offer...

...optimized for expression in E. coli: acoustic gene 1 (arg1) and acoustic gene 2 (arg2). The key difference... plasmids are available from Addgene! References 1 Bourdeau, R.W., Lee-Gosselin, A., Lakshmanan, A., ...have hollow interiors and a protein shell that excludes water but is gas permeable. These vesicles are...sequentially exposed to acoustic pulses with high enough amplitudes to first collapse arg2 and then arg1 vesicles...

.... pH and DNases affect DNA stability Figure 1: So many choices! What to do? The main factors ...your downstream experiment. TE (10 mM Tris-HCl,1 mM EDTA, pH 8.0) buffer is the best buffer for preserving... favors the breakage of the hydrogen bonds, destabilizing the molecule and favoring the separation of ...nuclease free water is a great option because it provides a great versatility on how to use your plasmid...

...while the paid version provides more tools for plasmid mapping and design (Figure 1). Benchling: While you...Davis from the University of Utah.   Figure 1: A plasmid map created in SnapGene of Blue Flame plasmid...primer design Design primers for Goldengate cloning   Design primers for In-Fusion Cloning Design primers...academic users. Serial Cloner: Free desktop-based software for plasmid design and mapping. ApE (A plasmid Editor...Primer design tools Many of the above-listed plasmid mapping tools have built-in primer design tools, ...stand-alone sites for general primer design, as well as primer design for different cloning applications...characteristics. NEB Data-optimized Assembly Design (DAD): This tool designs overhang sets for high complexity Golden...

...B. Gurumurthy, and Masato Ohtsuka. Nat Protoc. 13(1) (2018):195-215. PubMed PMID: 29266098 Creating knock-in..., with protocols using dsDNA templates achieving ~1-10% efficiency. Easi-CRISPR uses a long ssDNA template...working with CRISPR than just designing a repair template that contains your desired edit, and I want to make...say about CRISPOR, a tool to help you design, pick, and close guides for Cas9 and Cpf1. This tool supports...The blog post I mentioned, John Doench’s How to Design your gRNA for CRISPR Genome Editing, gets over ...these numbers show that this basic concept of gRNA design is still challenging for researchers using CRISPR..., and that standardizing and improving the gRNA design process and rules for various applications is necessary...

...takes a different approach, randomly removing 1-20 nucleotides from the cleavage site specified by the gDNA...About NgAgo User's First Impressions Here References 1. Gao, Feng, Xiao Z. Shen, Feng Jiang, Yongqiang Wu...DNA guides NgAgo uses 5’ phosphorylated DNA guides (so called “gDNAs”) rather than the RNA guides employed...24 base DNA guides may be more user-friendly, since they can be ordered as oligonucleotides and should ...gDNA guides will also need to be transfected into the cells of interest. In contrast, RNA guides must ...important role in RNA interference, binding RNA guides to cleave foreign RNA. Argonaute proteins are also...cleave DNA in vitro, directed by phosphorylated DNA guides. Unfortunately, Swarts et al. were unable to show...

...organoid model (Sun, et al. 2021).  Figure 1: Example schematic from the Organoid EasyTag system...without altering that of the internal control (Fig 1). Because this method relies on the fluorescence ratio...CRISPR inhibition (CRISPRi). A) Lentiviral vector design. B) Strategy for leak-free transcriptional control...Organoid EasyTag workflow combines thoughtfully designed plasmid components with Fluorescence Activated... heavy optimization. Claudia Vickers’s lab has designed novel SL fluorescent biosensors by integrating... rather than on the fluorescence intensity, it provides a direct and quantitative detection of SLs. The...biosensors could have a major economic impact.   Design and mechanism of action of the receptor-based SL...

...microscopic resolution for studying colloidal gold (1).  The method was based on using a thin plane (sheet...Siedentopf, and R,. Zsigmondy.  (1902) Ann. Phys.  315, 1 – 39.
 A. H. Voie, D. H. Burns, F. A. Spelman. (1993...microscopy/en_de/products/imaging-systems/lightsheet-z-1.html ...After this initial application, Stelzer’s group described the single-plane or selective-plane illumination...and acquire images of a sample from two opposing sides and merge the images, such as in the Ultramicroscope...

...determined LY6A, also known as stem cell antigen-1 (SCA-1), is the receptor used by PHP.B to reach the CNS...William L. Stanford. "Concise review: stem cell antigen‐1: expression, function, and enigma." Stem cells 25.6... a receptor for PHP.B. Read on to learn how the design of PHP.B may have contributed to this mouse-specific...Cre-dependent selection yields AAV variants for widespread gene transfer to the adult brain." Nature biotechnology...

...sarcoma-associated herpesvirus.       References 1. Snapp, Erik. "Design and use of fluorescent fusion proteins ...experiments or even negatively impact cells (see Figure 1). When behaving properly, FPs enable researchers ... disulfide bonds in the secretory pathway (Figure 1) [4,6–8]. Additionally, FPs that encode N-glycosylation... live cells in real time. Previously, we have described a number of practical considerations that should...

... spread out onto the plate. We test the (capped) 1:1 agarose/DMEM solution against a part of the body ...a 60 mm dish ready for picking is shown in figure 1.   Find Additional Useful Protocols on Addgene's Protocol...investigators to identify specific clones with desirable properties, either initially, or using secondary...

...everything I allow 1 slide for every 1 minute of talk length, and then I cut a few more slides. Do not run over...NatureJobs: Interviews: The All-Important Job Talk. 1. Watch other people give great talks We are not talking...suspense. Notice the simplicity of their slides and how the slides complement the spoken message. Learn from...speak to start creating slides or thinking about content. Create a Google.slides or Powerpoint file right...how these speakers talk without looking at their slides or notes. Listen to their stories and narratives...listeners to be thinking “What happens next?”  Describe how you pushed to a solution or overcame obstacles...term, but are worried that this might sound condescending, consider explaining the term on a slide but...

...guide RNA (sgRNA), provided it meets two conditions:  1) The sequence is unique compared to the rest of the...bystander bases” if you choose this system.  Fig. 1: Cytidine deamination takes place on the free strand...temperature-controlled genome editing. Nat. Commun. 2017 81 8, 1–9 (2017).Walton, R. T., Christie, K. A., Whittaker...vectors and design your sgRNA targeting sequence using one of the many freely available sgRNA design programs...many cases several exons’ worth of DNA. Any ∼20 nucleotides of DNA can serve as the target for Cas9’s single...Cas9 localizes the deaminase to where the edit is desired, and it performs the programmed edit within the...flexibility. A consideration with this tool is that the desired edit will be performed on all relevant bases within...

...than control, 1; very dark). Brighter fluorescence indicates a more successful gRNA design. Adapted from...In vitro screening for the best gRNA Figure 1: Validation of double strand break (DSB) mediated ...and greatest in scientific content.      References 1. Cho, Seung Woo, et al. "Analysis of off-target effects... screen them for your desired edit. Good news! Remember the primers you designed to generate your pCAG-EGXXFP...CRISPR/Cas9 system floating around, and many pre-designed plasmids containing these variants ready for purchase...that do it for you. One such option is the CRISPR Design tool supplied by the Zhang lab. This simple website... and cons of using many different types of gRNA design software. When you are happy with your selection...

... to as ssODNs (oligodeoxynucleotides)) are commonly used for smaller modifications (~1-50 base pairs)....Easi-CRISPR was shown to increasing editing efficiency from 1-10% with dsDNA to 25-50% with ssODNs in mouse models...ribonucleoproteins. Quadros RM, et al. Genome Biol. 18(1); (2017). PubMed.  Additional Resources on the Addgene...your desired edit are all you need to generate a knock-in.   General considerations for designing a repair..., and ultraviolet light. In this post, we will describe the mechanism of homology directed repair, one...increase the frequency of edits, researchers have designed self-cleaving plasmids that liberate the targeting...information on HDR and CRISPR. Our CRISPR 101 series was designed to help explain the basic principles driving CRISPRs...

...principles of fluorescent dyes/ligands Figure 1: Example structures of fluorophores, fluorescent dyes... dopamine D2/D3 receptors. Scientific Reports, 10(1), 21842. https://doi.org/10.1038/s41598-020-78827-...for bacterial imaging . Microbiology Spectrum, 12(1), e0369023. https://doi.org/10.1128/spectrum.03690...Biochimica et Biophysica Acta - Biomembranes, 1838(1), 15–33. https://doi.org/10.1016/j.bbamem.2013.09.005...select the optimal dye or fluorescent ligand when designing an experiment. You may find yourself defaulting...protein, enabling the labeling of protein substrates/peptides and antibodies. Example applications of this include..., which is an important consideration for assay design (Breen et al., 2016). When selecting dyes, it's...

...catalog had grown to 879 viral catalog items (Figure 1). The service caught on quickly in the scientific ...over 100,000 viral vector preps!   Figure 1: A) Cumulative AAV preps available in the Addgene ...Neuromethods, 195, 77–101. https://doi.org/10.1007/978-1-0716-2918-5_5 Haery, L., Deverman, B. E., Matho, K...site, and titer. Addgene’s Viral Vector Service Besides being complex to use, viral vectors are also difficult...which include calcium and voltage sensors, are the broadest category of viral vector tools, which may help... for vectors in the “controls” category, which includes things like viral vectors that deliver GFP. ...most requested serotypes (Figure 4), our catalog includes vectors in a variety of serotypes, including the...

...your budget, you should take the following steps: 1. Define your timeline Start with your thesis in mind...these decisions in the following order of importance: 1. First make decisions based on experimental timelines... several factors aren’t taken into consideration. 1. Always focus on the science When negotiating finances...Wilpitz founder and lead consutant at Experiment Designs Consulting, Inc. “How much do you need [for your...necessary resources. This is why at Experimental Designs Consulting (EDC), we instead have our clients focus...the institution/funding source. Major equipment includes large centrifuges, sequencers and analyzers, and... as many of these tools as possible, this even includes clocks, timers, and phones.  As for reagents and...

...protein sequence of the predecessor GEVIs (called ASAP 1 and 2), they found an indicator that is optimized ...several other constructs in plasmid form. Fig. 1: Summary of the approach and outcomes of the screen...optimized for 2-photon imaging of voltages. This includes monitoring of fast bursts, as well as slow up/down voltage...KCRs), from a fungus-like protist Hyphochytrium catenoides (Govorunova et al. 2022). The authors show that...

... similar antibodies in llamas and alpacas (Figure 1). At the time, researchers were already on a quest...nanobodies (Arbabi-Ghahroudi, 2017)!      Figure 1: Comparison of a heavy-chain only antibody and single-domain...camelids too, though not just for the reasons the UN describes. In honor of the Camelid family, I wanted to take...of times on this blog over the years and have described how they compare to other antibodies and affinity...

...into E. coli is a fairly inefficient process– just 1 out of 10,000 cells on average! Without some means...down especially fast and plates should be used witin 1 month for optimal efficiency. Beware of satellite ...when the cells are grown on selective media, and provides those bacteria with a pressure to keep your plasmid...gene and acts as a marker for the mutation. When designing these types of experiments it is best practice...

...and drawbacks of four split AAV vector approaches. 1. Overlapping  Overview: The overlapping strategy has...Adeno-Associated Virus Capsids. Human Gene Therapy Methods 27:1–12 . https://doi.org/10.1089/hgtb.2015.140 Frischmeyer...doi.org/10.1002/emmm.201302948 Trapani I, Toriello E, de Simone S, Colella P, Iodice C, Polishchuk EV, Sommella...physical size of the genomes is still ~4.7kb. But despite this, larger functional transgene products were...HDR event can’t occur. Lastly, depending on its design, donor A may be capable of expressing a truncated...

...P2A-PuroR for lentiviral expression. Figure 1: dCas9-p300 adds H3K27ac marks to histones in promoter...lentiviral transduction. Lysine-specific Demethylase 1 (LSD1) Tatjana Sauka-Spengler's lab has deposited ...oncogenic silencing in vivo by programmable and targeted de novo DNA methylation in breast cancer. Oncogene 34...which uses light to regulate transcription, also includes light-regulated histone methyltransferases and...histone modifications. Like the TALE-LSD1 system described above, dCas9-LSD1 inactivates targeted enhancers...certainly not the only CRISPR-based technology designed to alter gene expression. Fusing dCas9 to a transcriptional...epigenetic modifiers? As with any experiment, your desired outcome will determine the tool that you should...

...et al., 2019; McLaughlin, 2019).    Figure 1: Schematic of multiplex immunofluorescence approaches... Brewer, M., McDonough, L., Zhu, Y., Neumann, E., De Caestecker, M., Gutierrez, D., & Spraggins, J. (2022...). Springer New York. https://doi.org/10.1007/978-1-4939-8935-5_26 Jambor, H., Antonietti, A., Alicea,...between the targets. Or, it might be sufficient to describe or classify the labeled structures visually; for...will vary depending on your specific experimental design. Like selecting an antibody, some of the requirements...

...Full "Applying for a Faculty Position" eBook Here 1. Have a competitive CV You don't necessarily need ...that people would agree is important to understand. (1-2 sentences). For example: Distinguishing correctly... What will be the focus of your first grant? (1-2 sentences) Simply stated, what are your aims...Applying for a Faculty Position: the View from Both Sides," are summarized below. Check Out Erik Snapp's ...You want people to focus on you and your ideas. Slides will be     distracting. What is the big question...feedback. Many of these documents are too vaguely described in other sources of guidance or the document examples...single malt whiskies, but I remember the days of modest postdoc salaries all too well. If there's a group...

...sequences were initially discovered in E. coli in 1987 (1), the concept that these clustered repeat sequences...neutralize CRISPR/Cas. Earlier this year, expression of 1 of 5 anti-CRISPR genes were found to inactivate the...multiplexibility in various species and describe an assortment of applications. Despite the ubiquity of CRISPR/Cas ... store a record of invading phages but to also destroy the phage upon re-exposure (2, 3). More specifically...complex binds to the foreign DNA, a cut is made to destroy the invader. Wikipedia, accessed 25 November...engineering. The diversity of the CRISPR/Cas systems provides powerful lines of defense against invading phages...CRISPR/Cas inspired genome editing was initially described by Cong et al. and Mali et al. in January 2013...

...has error rates that range from roughly 1 per 500bp to roughly 1 per 10 million bp depending on the polymerase...for Gibson assembly. The steps following primer design and the PCR process itself are very similar to ...the downstream steps. For this example, we will describe how to copy a cDNA from one vector into a new ...YGOI) for ligation into a recipient plasmid. Designing primers for PCR based cloning     The basic...: Do not cut within your insert Are in the desired location in your recipient plasmid (usually in ...examine the DNA sequence that we want to amplify and design primers that will bind to and replicate it. The...ends of the ORF and how these are used for primer design: Because we are cloning an ORF, we want to clone...