...Resources on the Addgene Blog Learn to do Restriction Cloning Try SLIC cloning Learn about Gibson Assembly ... One key difference is the plasmid DNA must be released from the bacteria in order to serve as PCR template...reaction. You just need to lyse the bacteria to release the plasmid DNA by either briefly boiling the sample...Read about Choosing a Molecular Cloning Technique Learn to Perform a Bacterial Transformation Deposit Your...

...visualizing promoter activity, and much more. Read on to learn more about GFP, how scientists have evolved this...exist with only slight variations separating them.  Please note that many fluorescent proteins found on the...F64L; S65T, Y66W; N146I; M153T; V163A; A206K Cerulean F64L, S65T, Y66W, S72A, Y145A, H148D, N149I, M153T... Y145A and H148D Stabilizes the structure of Cerulean derivatives F99S, M153T, V163A Improves folding...visualize the effects of alterations to this activity. Learn more about optogenetics at OpenOptogenetics and ...

...remains unclear whether the FP misfolding impacts fusion protein function and this could lead to under...localize [4]. Most investigators are surprised to learn that FPs are often poorly suited for cellular compartments...glycosylation, disulfide bond formation, and proteolytic cleavage. FPs localized to organelles are equally susceptible... dark fluorescent proteins in your experiments, please consider the following: How can one ensure that...

...bit of a steep learning curve and many opportunities for mistakes. The easiest way to learn optogenetics... is an approximation of reward or the sense of pleasure that the animal perceives through taking a drug...University.  His interests include the role of learning and memory in opioid addiction and science communication...Kheirbek, Mazen A., et al. "Differential control of learning and anxiety along the dorsoventral axis of the...

...CROPseq-multi using selection by Puromycin, Zeocin®, or nuclear expression of BFP2.     Figure 1: CROPseq-multi...separated multiple sgRNAs with tRNAs, which are cleaved out of the RNA transcript by endogenous processing...and found the protein La (a small RNA-binding exonuclease protection factor) promotes editing (Yan, J et...hypothesized that the improved efficiency is at least in part due to La binding and stabilizing the 3’...Neurodegeneration Collection updates Last but not least, check out the new and noteworthy tools featured...

...each viral lot. To learn which specific QC experiments were performed on your lot, please contact us. Full...medium (OptiPro SFM) containing lentivirus is first cleared by low-speed centrifugation and then by filtration...responsive element (RRE) gene and the cellular ribonuclease P/MRP 30 kDa subunit (RPP30) as a control for...the sample from the DNA tube all the way to the cleared viral preparation pool. In addition, the final ...

...integrating neuronal signalling and triggering release of the stress hormone norepinephrine to promote...are chemogenetically activated, norepinephrine release causes a freezing response, a form of immobility... the NAc, where enhanced glutamate transmission leads to locomotion sensitization. (for review see van... as sleep, food intake, mobility/coordination, learning, and memory. We’ll cover these in a  future blog...Guide to Viral Vectors Resources on Addgene.org Learn about Addgene's viral service Find viral protocols...

...tips from an AAV rookie  Learn about some considerations when using AAVs Learn more about using AAV as ...complex (LY6A) as a receptor for PHP.B. Read on to learn how the design of PHP.B may have contributed to ...182-188. PubMed PMID: 29175632. If you want to learn more about how PHP.B and PHP.eB were engineered,...PMC5088052. Additional resources on the Addgene blog Learn more about PHP.eB, and PHP.S, a peripheral nervous...

...Blog Learn All about Mammalian Vectors Read up on CRISPR Delivery Systems for Mammalian Cells Learn about...older than those available from biorepositories leading to genotypic and phenotypic artefacts. The life...commercial repositories and the same cell line from a colleague?”. The main difference is the QC and validation...usually the case if you obtain a cell line from a colleague. Time and money are wasted, papers are retracted...

...replicon of pTargetF may exhibit a more serious leaky expression in BL21(DE3), which we presume caused...that indeed the gRNA-pMB1 had a slightly higher leaky expression in BL21(DE3) that resulted in a lower...a sacB gene renders it beneficial for plasmid clearance. It is more convenient to update the spacer(20nt...Anhydrotetracycline (aTc) induces expression transposases leading to increased integration frequency. (C) Increasing...rounds of transfer on transposition-inducing agar lead to higher cargo copy numbers. (D) The effect of ...

...different chromatin states. Alexander Stark and colleagues at the Research Institute of Molecular Pathology...Initiative, led by Dominic Esposito, has recently released a collection of 360 Ras pathway plasmids to the...Drosophila cells, targets 13,501 fly genes with at least three independent gRNAs, each chosen strategically...vacuolar) pH environments that inactivate SEP and leave only DsRed to fluoresce (1). With variants that ...effects on gene regulation. Professor Zoldos and colleagues from the Department of the Biology, Division ...26912792 FusX-TALEN assembly system TALEs as nucleases (TALENs) are genome-editing tools that are deployed...auxotrophies or using different auxotrophic backgrounds can lead to disparate physiological effects and complicate...

...Bench series was born from the shift to remote learning due to COVID-19. We had many protocols for plasmid...details about personal protective equipment to learning about how to use a tissue culture hood and other...other lab equipment, we’re here for you as you learn the ropes! All material can be found on our Protocols...Protocols page and YouTube page.  Happy learning, and welcome to the lab bench, new scientists!   ...

...warm meal on a winter day, you should be happy to learn that many DNA-based reagents are named for soup ... and non toxic to cells. Because it rapidly photobleaches, DFHBI can only be used for qualitative, but...regions in the 5’ UTR of mRNA that bind metabolites, leading to conformational changes that control the expression...the Corn-DFHO complex. For dessert (last but not least), the only fruit-named aptamer, Mango, binds to ...PMC4143336. Additional resources on the Addgene blog Learn more about aptamer fluorophores Investigate other...

...the uses of biotinylation in molecular biology! Learn about in vivo biotinylation of bacterial fusion ...biotin-avidin complexes bound to your protein of interest leading to amplification of the reporter signal and easier...microenvironments. There are also reversible and cleavable biotinylation reagents to help with the specific...vivo biotinylation of bacterial fusion proteins Learn how biotin is used in Split-BioID: An improved method...method for studying protein-protein interactions Learn how biotin can be used with CRISPR Additional resources...

...expressed proteins endA, endA1 Endonuclease I (nonspecific cleavage of dsDNA ) mutation Improves plasmid...cell mcrA, mcrBC Inactivation of pathway that cleaves methylated cytosine DNA Allows for uptake of foreign... mrr, Δ(mcrC-mrr) Inactivation of pathway that cleaves methylated adenine or cytosine DNA Allows for uptake...recombination, increases plasmid stability recBCD Exonuclease V activity abolished Increased plasmid stability...

...blog posts Learn more about AAVs Resources on the Addgene website Visit the AAV Data Hub Learn more about... serotype, injection site, and incubation time. Learn more about the Data Hub in this blog post. Alongside...attending SfN2019, stop by booth #2163 to say hi and learn more about Addgene’s viral service. Additional ...

... the Addgene Blog Learn How Piggy Bac Vectors Can Be Used for Genome Editing Learn about Michael Koeris...needs. For more context, exposition, and detail, please refer to the paper. Why might one be interested...uploaded to Addgene. We also recommend inserting self-cleaving hammerhead ribozymes directly upstream of the ... transcribed gene and its 5’ UTR. Various self-cleaving hammerhead ribozyme parts are described in Nielsen...

... I had found my dream job, because one thing I learned in grad school was that scientists have amazing...at their lab benches. It’s these exchanges with leading scientists (and our growing collection of 25,000...sciences research for the past decade. We’ve also learned along the way that a connected and collaborative...scientists who have deposited plasmids at Addgene.  Learn how discoveries were made and get practical tips...

...Health and Allied Sciences in Ghana. Read on to learn more about doctors Bengyella and Duedu and how they...tumors is “a major public health problem, and a leading cause of hospitalization for gynaecologic disorders...Program Additional Resources on the Addgene Blog Learn about Plasmid Awards to Researchers Working on Rare... in China Additional Resources on Addgene.org Learn About Addgene's Mission Check Out Our International...

...even to not bind to their target molecule at all, leading to inconclusive results, erroneous conclusions,... that your antibody has been validated using at least one of these strategies. Choosing which strategy...) the protein of interest in your sample, should lead to reduced or eliminated antibody signal. If you...translation start sites or alternative splicing that could lead to some form of the target protein to be present... Check out our Selecting an Antibody blog post! Learn more about Antibodies and their many applications...