We narrowed to 628 results for: Ncl
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TypeBlog PostPublishedSept. 27, 2018, 1:09 p.m....completely understood, but some possible reasons include: Metabolic pressures - it takes cellular resources...when designing a full gene. A non-exhaustive list includes: Codon abundance relative to cognate tRNA abundance...insoluble, nonfunctional globs of protein known as inclusion bodies that will generally segregate with the ...assay to make sure your protein isn’t forming inclusion bodies and is folding properly. References 1...
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Changing Labor Laws Bring Increased Postdoc Wages
TypeBlog PostPublishedOct. 25, 2016, 2:30 p.m....administered, which leads to many complications, including the inability of some institutions to know exactly...Furthermore, 7% did not enforce these policies. It is unclear how many postdocs are at each of these institutions...are afoot for the postdoc population, and it is unclear how this will all shake out. In the short term,...would help our resource. In the medium term, it is unclear what effects the new laws will have on the postdoc... -
Important Considerations in Optogenetics Behavioral Experiments
TypeBlog PostPublishedOct. 1, 2015, 2:30 p.m....but some examples I’m personally interested in include: intracranial self-stimulation (ICSS) and place...resolve these issues is with electrophysiology. Concluding thoughts and moving beyond the mouse While it...applicable to the rat [6]. Technical differences include the ferrule implants and patch cables. One huge... at the Rockefeller University. His interests include the role of learning and memory in opioid addiction... -
Antibodies 101: Introduction to Immunofluorescence
TypeBlog PostPublishedNov. 16, 2021, 2:15 p.m....depend on target and sample type. Example steps include: permeabilization, antigen retrieval, and sectioning...available, and your experimental goals. Example steps include: counterstaining, mounting sample on slides, and...you optimize your protocol, always remember to include controls to show that your antibodies are binding...antibodies validate them in a variety of applications, including IF (or IHC or ICC), and will share some of their... -
Fluorescent Proteins 101: When GFP lets you down
TypeBlog PostPublishedAug. 23, 2018, 12:05 p.m....Several factors contribute to the popularity of GFP including (i) fast and complete maturation to functional...endogenous fluorescent co-factors can be used. Examples include the LOV domains that bind flavin (Buckley et al... conditions, with pKa values below 4. Examples include mTurquoise2 (pKa = 3.1), tagRFP (pKa = 3.1) and... or media. These sources of ‘autofluorescence’ include riboflavin in the medium or flavin containing proteins... -
Viral Vectors 101: AAV Serotypes and Tissue Tropism
TypeBlog PostPublishedJan. 16, 2025, 2:15 p.m....AAVs to broadly spread throughout an organism, including difficult-to-target tissues like the central nervous...and so on, although we often use abbreviated nomenclature (AAV1, AAV2, etc.) to distinguish the capsids...AAV pseudotyping. It has broad tissue tropism, including liver, muscle, lung, CNS, kidney, retinal, and...serotypes used to target the central nervous system, including neurons, AAV5 is generally the best choice for... -
RNA Interference in Plant Biology: New Tools for an Old Favorite
TypeBlog PostPublishedOct. 27, 2020, 1:15 p.m....afflicted by pervasive off-target effects. This conclusion was based upon numerous phenotype discrepancies...importance of using orthogonal methods to confidently conclude the function of a gene of interest. Tools for ...discussed below). Examples of these constructs include pGAPi, which allows easy creation of long hpRNA...experiment are now available on Addgene. These include the construct that allows for easy insertion of... -
Mycoplasma Contamination: Where Does It Come From and How to Prevent It
TypeBlog PostPublishedOct. 8, 2020, 1:15 p.m....various sources that are difficult to trace. These include the laboratory personnel, the serum, the cell culture...cross-contamination and due to poor lab techniques. These include reusing pipettes for multiple cell lines rather...Contamination! Tissue culture media often have indicators included in the media that turns yellow from lowered pH... periods and affect the total cell viability. Conclusion To sum up, it is always important to prevent ... -
Finding Your Science Policy Path
TypeBlog PostPublishedSept. 22, 2020, 1:15 p.m....organizations in a variety of science policy roles including fellowships, volunteer positions, and part-time... with a large set of other societal pressures including economic, cultural, and political. Science policy...science blogs. Other avenues for skill development include leadership of extracurricular organizations, working...opportunities. NSPN maintains a great resource guide including conferences, societies, and outreach groups. Grab... -
Antibodies 101: Validation
TypeBlog PostPublishedMarch 24, 2022, 1:15 p.m.... molecule at all, leading to inconclusive results, erroneous conclusions, and wasted time. In fact, many...antibody-dependent approach. Antibody-independent methods may include analysis of transcriptomic or proteomic data from...specifically to the expected protein. Common approaches include using RNAi to knock-down expression of your target... -
Sequencing Options for CRISPR Genotyping
TypeBlog PostPublishedOct. 4, 2016, 2:30 p.m....for CRISPR genotyping. The most common options include mismatch cleavage assays, such as Surveyor™, T7E1...information profile regarding putative off-target sites including both the edit rate and the repair product sequence...algorithms do not take into account other elements, including those relating to DNA structure (e.g. epigenetic...options exist for unbiased off-target detection, including Digenome-seq (Kim et al. 2015) for in vitro analysis... -
Cancer and the Immune System: Deciphering the Relationship
TypeBlog PostPublishedMarch 14, 2017, 2:30 p.m...., introduced the “Three Es of Immunoediting” nomenclature. 1. Eliminate - Latent tumors are recognized...on immune system Conventional cancer therapies include radiation therapy (owing to contributions from ...blocking. Other molecular targets under research include TIM-3, LAG-3, STAT-3, EGFR, CD20 and CD73 on specific... factors researchers are beginning to focus on include: Cell heterogeneity Tumors are composed of a wide... -
Antibodies 101: Introduction to Antibodies
TypeBlog PostPublishedJan. 19, 2021, 2:15 p.m.... Figure 1: Labeled diagram of an antibody including Fc, Fab, heavy chain, light chain, constant region...antibodies for experiments in a variety of fields including neuroscience and immunology. They can be used ...analyzed using techniques such as flow cytometry. Conclusion Whew! Now you know that antibodies can be used...used for. Addgene’s Antibody Plasmid Collection includes plasmids that express antibodies, nanobodies, ... -
CRISPR 101: Multiplex Expression of gRNAs
TypeBlog PostPublishedSept. 10, 2020, 11:45 a.m....for plasmids suitable for other model systems, including E. coli, plants, Drosophila, and zebrafish! ...destination vectors. Destination vector options include humanized wt Cas9, dCas9 (transcriptional repressor... Figure 3: Comparison of Multiplex Strategies including Standard PolIII-gRNA cassette, Csy4-cleavable ...filler sequences. You also have the option of including a previously-designed tyr gRNA, which causes hypopigmentation... -
Selecting Your Plasmid Purification Kit
TypeBlog PostPublishedMay 27, 2025, 1:15 p.m....sneak in from almost any step of the process and include things like genomic DNA, RNA, proteins, salts, ...sustainable practices for the lab. Plastic consumables (including pipette tips, plates, and yes those purification... kits from a variety of manufacturers. It also includes a rough comparison of list price between these...Yeast, BAC, transfection-grade While we’ve included a sample of some commonly used kits, please note... -
Plasmids 101: How to Name Your Plasmid in 3 Easy Steps
TypeBlog PostPublishedFeb. 13, 2014, 1 p.m....for plasmid. pXXXXX-XXXX Step 1: Backbone name Include the empty backbone name in your plasmid name. This...backbones, visit Vector Database. Step 2: Insert name Include information about the insert in your plasmid name...contains a mutation or modification, this should be included in the plasmid name. Mutations are generally listed... -
Prime Editing: Adding Precision and Flexibility to CRISPR Editing
TypeBlog PostPublishedJan. 13, 2025, 9:49 p.m....the template. The PE3 system does just this by including an additional sgRNA. Using this sgRNA, the prime... number of unintended prime editing outcomes, including indels. Therefore, they engineered a dominant-... ✓ ✓ reflects an innovation always included in the indicated tool. ~ reflects an innovation... strategies in our CRISPR guide. Some examples include: TwinPE: two prime editors make complementary ... -
The Addgene Antibody Data Hub
TypeBlog PostPublishedSept. 12, 2024, 1:15 p.m....species, antibody species, or whether the results include knockout data. Figure 2: Search options in ...description of the experiment. If that data was included in a publication, you'll find that information... is easy! Here at Addgene, we love all data, including negative data, which can help researchers save... -
Making Career Development an Integral Part of the Science Training System
TypeBlog PostPublishedFeb. 25, 2021, 2:15 p.m.... to a post-academic career. This training was “included” with their tuition, so to speak. Unfortunately...the development of priorities for pd|hub which include building capacity to identify the most effective...scientists, but it is essential that we integrate inclusive career readiness training in this educational ... -
What's New in CRISPR - May 2019
TypeBlog PostPublishedMay 28, 2019, 12:49 p.m....per gene and includes ~10,000 gRNAs targeting intergenic regions. The libraries also include 2,400 gRNAs...recently deposited to Addgene and you’d like it to be included in the next What’s New in CRISPR blog post, please...