We narrowed to 910 results for: ire;
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TypeBlog Post...with multiple inserts or internal mutations that require more than two sequencing reactions. In the more...to ensure uniformity of the culture. Sequencing directly from the glycerol stock saves us the step of purifying... Each sequencing service will have different requirements for volume and concentration, and not all providers...
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Career Coaching for Scientists: Why and Where Do I Find One?
TypeBlog Post...new job skill? Finish your thesis? Knowing your desired outcomes will help you assess if the coach you ...coach usually will be a clear communicator and make direct suggestions for improvement and change. Work ...relationship has been successful – what are the desired outcomes? Ask openly for clear information on... -
Plasmids 101: Cre-lox
TypeBlog Post...asymmetry in the core sequence gives the loxP site directionality, and the canonical loxP sequence is ATAACTTCGTATA-GCATACAT-TATACGAAGTTAT...reversed. Deletion: If the sites face in the same direction, the sequence between the loxP sites is excised...cerevisiae FLP-FRT recombination system is another site-directed recombination technology very conceptually similar... -
Plasmids 101: The Wide World of Natural Plasmids
TypeBlog Post...viral infection by bacteriophages), or conjugation (direct transfer from one cell to another). Bacteria need...transfer genetic material to a recipient through direct contact. That’s conjugation! F factors can also...cloning. In addition, scientists have engineered entirely new types of plasmids without clear analogues ... -
Plasmids 101: Modular Cloning
TypeBlog Post...simple, and engineering complex cellular behaviors requires making complex manipulations to cells. For example...number in the Level 2 assembly. These pathways can require a lot of optimization - which is why MoClo is so...of a combination of multiple genes to achieve a desired effect. This is commonly used in plant biology ... -
Viral Vectors 101: Chemogenetics
TypeBlog Post...Targeting Receptor Expression Using chemogenetics requires successfully introducing receptors to target cells... to control ion flux. The system allows you to directly manipulate ion channels. When working with PSAM...Magnus C, Lee P, Bonaventura J, Zemla R, Gomez J, Ramirez M, Hu X, Galvan A, Basu J, Michaelides M, Sternson... -
Getting Started with Behavioral Neuroscience: Standardizing Experiments and Using Viral Vectors
TypeBlog Post...laboratory, a high level of experimental control is required, because seemingly tiny external factors can have...overexpress a gene that alters neuronal function directly such as receptor subunits or neurotrophic factors...smaller and may be preferable if wider diffusion is required. On the other hand, lentiviral vectors are superior... -
Open Resources and Plasmid Tools For Studying C. elegans
TypeBlog Post...Development Dynamics Database (WDDD) The WDDD aims to acquire quantitative data on all the cell divisions of ...Kit for CRISPR Cas9 edits At Addgene, we have the entire SapTrap toolkit from the Jorgensen lab. SapTrap...Project: The Complete Gene Expression Map of an Entire Nervous System." Neuron 99.3 (2018): 430-433. PubMed... -
Hot Plasmids and Viral Preps - January 2021
TypeBlog Post...active sites. In addition, Casɸ has a minimal PAM requirement. For instance, for Casɸ-2 the PAM sequence is... is expected to contribute to experiments that require high photostability, such as time-lapse imaging...Leopold Parts. This library delivers 2 randomly paired guides per construct and allows a reduced scale... -
Split Fluorescent Proteins for Studying Protein-Protein Interactions
TypeBlog Post..., comprised of PAS and GAF domains of iRFP713. Requires presence of heme oxygenase for chromophore formation...miRFP709 and miRFP670, pair with common PAS domain. Requires presence of heme oxygenase for chromophore formation...BiFC interaction is typically irreversible and requires some time for chromophore maturation, splitFAST... -
CRISPR in the Clinic
TypeBlog Post...Systemic therapies involve CRISPR-Cas delivery directly to the patient. Patient-derived cells can alternatively...the goal of ‘fixing’ mutations or introducing a desired edit, but this particular class of treatments has...inflammatory diseases, blindness, and diabetes. Directions and obstacles for future CRISPR therapies CRISPR... -
High Complexity Golden Gate Assembly with NEB’s DAD
TypeBlog Post...longer incubation times and didn’t necessarily require cycling. The Pros and Cons Researchers used the...select for correctly assembled phage genomes, it required a very stringent selection process and benefited...selection, failed under normal cycling protocols, and required a 48-hour incubation at 37 °C. While the 52-piece... -
Fluorescent Proteins 101: Introduction to FRET
TypeBlog Post...photon 2. Energy transfer from the excited donor directly to the acceptor fluorophore--think of it as a ...fusions varies with transfection efficiency and any unpaired fluorescent proteins can contribute additional...Check Out Fluorescent Protein Biosensors Browse the Entire Fluorescent Protein Collection ... -
Plasmids for Endogenous Gene Tagging in Human Cells
TypeBlog Post...effectively utilize the host cell’s inherent homology directed repair (HDR) process following the double strand...break and introducing the tag through homology directed repair To introduce the fluorescent tag to the...on the Addgene Blog Learn More about Homology Directed Repair Learn More about Non-Homologous End Joining... -
Plasmids 101: FLEx Vectors
TypeBlog Post...that the orientation of the flanking loxP sites directs the Cre recombinase to invert, translocate, or ... the antisense orientation (Figure 1, top). The entire DNA cassette would be flanked by two pairs of target... Wendling O, Chambon P, and Ghyselinck NB. A directional strategy for monitoring Cre-mediated recombination... -
Reaching out to China: Canton Nucleic Acids Forum (CNAF) 2015
TypeBlog Post...world. With three Nobel Prize winners, one NIH Director and a co-founder of BGI-China presenting, it’s... discoveries and moving beyond DNA Eric Green, Director of the National Human Genome Research Institute...proteins). The chemistry is complex and this work requires an interdisciplinary team. Day two of the conference... -
Magnetic Control of Proteins: More than a Dream
TypeBlog Post...deposited their plasmid with Addgene, hoping to inspire others to explore the phenomenon, improve the tool...Evolution of a new variant Maria and the team set up a directed evolution and screening experiment with three ...topics. He embraces open science, self-publishing directly on his website (and often posting updates on social... -
Antibodies 101: Flow Compensation
TypeBlog Post...This will make it difficult to properly gate your desired cell population. By compensating between fluorescent...other corners of the dot plot, compensation is not required. In Figure 3, we also know that on most cytometers... cytometers, FITC and APC should not require compensation between each other. We can therefore conclude... -
Hot Plasmids and Viral Preps - May 2021
TypeBlog Post...syndrome coronavirus 2 (SARS-CoV-2), the virus may acquire mutations to escape these therapeutics. Jesse Bloom...the antibodies were able to immunoprecipitate the entire complex in three different cell lines. The Gräslund...enzyme. Compared with previous research, which required protein compartments to co-localize multiple enzymes... -
Adenoviral Delivery of CRISPR/Cas9 Aims to Expand Genome Editing to Primary Cells
TypeBlog Post...alternative approach to NHEJ is to utilize the Homology Directed Repair (HDR) to knock in corrective genes. For...to repair the double strand break via homology directed repair and you would probably need to use a two...human cells being subjected to scarless homology-directed genome editing (Maggio et al., 2014). Gonçalves... -
Multiplexed Capture of Promoter-enhancer 3D Chromatin Structures Using CRISPR
TypeBlog Post...3C-based sequencing requires a large number of cells (~5x107). Together, these two requirements prevent the use...original CAPTURE method The original CAPTURE method requires the creation of a stable cell line that co-expresses... -
Transferable Skills Guide: Managing a Team
TypeBlog Post...handle or direct with a degree of skill, To work upon or try to alter for a purpose and To direct the professional... these tasks before I was a manager! Second, my direct report was not engaged because I only left him/... -
CrispyCrunch: High-throughput Design and Analysis of CRISPR+HDR Experiments
TypeBlog Post...single 96-well plate for HDR that satisfy all requirements. To alleviate this bottleneck, we began building... your own purposes, or delete the originals if desired. It works as a whole or in part. For example, you... were optimized for HDR, except for TagIn. HDR requires the additional step of mutating guide sequences... -
Starter guide to induced pluripotent stem cells (iPSCs) part 1: A renaissance in regenerative medicine
TypeBlog Post...ethical concerns of ESCs. As iPSCs can be sourced directly from a patient’s adult cells when used therapeutically... are low. Further, as iPSC generation does not require embryos, their use skirts issues surrounding the...formation, commonly referred to as reprogramming and, direct conversion from one somatic cell type to another... -
Golden Gate Assembly Upgrades: More Fragments, Faster Assembly, and Higher Fidelity
TypeBlog Post...yields from entire outgrowth built from 2 µl of the assembly reaction, and from the entire assembly reaction...assemblies) translate to faster assembly workflows that require less screening to confirm the accuracy of assembly... -
Viral Vectors 101: Voltage Indicators
TypeBlog Post...level. The heartbeat – a muscle contraction – is directly related to the electrical impulse of the cardiac... to measure membrane potential and can also be directed to specific types of cells, such as neuron subtypes...For in vivo imaging applications, the ability to direct your voltage sensor selectively is especially essential... -
AAV Vector Quality Control: Going the Extra Mile with NGS
TypeBlog Post... minor pre-recombined population that does not require the recombinase for transgene expression. Once ... of quality control is recommended but not yet required for clinical-grade materials, and is not commonly...https://doi.org/10.1089/hum.2019.277 Lecomte E, Tournaire B, Cogné B, Dupont J-B, Lindenbaum P, Martin-Fontaine... -
Cloning Mammalian Cells with the Agarose Method
TypeBlog Post...of antibiotic-resistant cells; during the time required to grease, place and fill a dozen cloning rings...endocrine cell lines we most commonly work with, require extra serum to survive at low densities - adding...growing cells such as HEK and CHO do not seem to require this, while nearly all endocrine cells do. While... -
Addgene's Cancer Collection Pages Connect the Research Community
TypeBlog Post...suppressors and upregulation of proto-oncogenes can directly inhibit the ability of a cell to undergo programmed...that a number of novel small molecules can bind directly, albeit weakly, to K-Ras and inhibit its activation...A single mouse click on a gene of interest will direct the investigator to a list of all available Addgene... -
Experimenting with New Careers while in Grad School
TypeBlog Post...guest blogger Melanie Fox, founder and executive director of Central Indiana Science Outreach and a Postdoc...as a student. They didn’t always know the best direction to guide me because they had no experience with...Melanie Fox is currently the founder and executive director of Central Indiana Science Outreach and a Postdoc...