We narrowed to 997 results for: lars

...Goedhart and Marieke Mastop from the Section of Molecular Cytology and Van Leeuwenhoek Centre for Advanced... can be excited many times, thereby producing a large number of emitted photons before they are destroyed...absorb the excitation light less efficiently. Similarly, decreasing the power of the excitation light ...these factors, environmental conditions such as cellular redox state and oxygen concentration may affect...Goedhart is an assistant professor at the Section of Molecular Cytology and Van Leeuwenhoek Centre for Advanced...Marieke Mastop is a PhD candidate at the Section of Molecular Cytology (University of Amsterdam) where she develops...Backbones with Fluorescent Tags Find Plasmids for Subcellular Localization Browse the entire Fluorescent Protein...

...experiments ranging from transient gene expression in a particular cell type to CRISPR genome editing, optogenetics... also be used to ensure specificity; this is particularly helpful when wanting to target specific neuron...vector. Full-length ssAAV vector genome of both polarities are rescued from the vector plasmid and individually... conversion of the DNA goes through the inter-molecular annealing or second strand synthesis. In contrast...transcriptionally active double-stranded form through intra-molecular annealing. Figure and legend from Okada, 2013....Production System: Towards Gene Therapy For Duchenne Muscular Dystrophy. InTech. doi: 10.5772/53023 Veldwijk...method for the titration of AAV-2 vector stocks."Molecular Therapy 6.2 (2002): 272. PubMed PMID: 12349826...

...difficulties in amplifying a template, particularly from a large genome, you can recheck your primer design...plasmid cloning Read our molecular biology reference Find how-to videos for molecular biology and lab techniques...sequence, or if you want to amplify sequences from a large genome organisms like mouse or human. There are ...nanodrop to assemble the  fragments based on their molar ratio. Assembly of the fragments There are multiple...the backbone plasmid; a starting point could be a molar ratio of 1:2 (plasmid:insert), but this parameter...

...two are present; intramolecular events happen with greater frequency than intermolecular events, but any... kDa protein responsible for intra- and inter-molecular recombination at the loxP recognition sites. A...sequence between the loxP sites is excised as a circular piece of DNA (and is not maintained).  Translocation... is excised, and gene expression proceeds. One popular lox-stop-lox plasmid is from Tyler Jack's lab: ...purposes, including activating an oncogene only in a particular organ, or bypassing embryonic lethality. To ...site-directed recombination technology very conceptually similar to Cre-lox, with flippase (Flp) and the short flippase...

...Common self-labeling tags Let’s take a look at some popular self-labeling tools: Tetracysteine: The first reported...variants run up to HaloTag11. HaloTag is now the most popular self-labeling tag in Addgene’s collection. CLIP-tag...on the protein of interest. Table 1: Summary of popular self-labeling tag systems Tag Size (amino... HaloTag for their "ticker tape" method for intracellular recordings, switching between different colored...Cornish, 2011). HaloTag and SNAP-tag are the most popular self-labeling enzymes in Addgene’s collection, ...that self-labeling protein tags are increasingly popular. Embrace the flexibility and switch things up for.... Two-colour live-cell nanoscale imaging of intracellular targets. Nature Communications, 7(1), Article...

...pitfalls of accepting alternative facts. Because, similar to scientists, the legacy of each government and...scientific consensus and calling for billions of dollars in cuts to important research funding.  So when...sacrifice in April because devaluing science has alarming implications for our public policies—many of which...that the community of science supporters is much larger and stronger that I expected. Jennifer Stancill...Birmingham, AL, USA 33.5207°N 86.8025°W - Jonathan Aguilar Galileo wrote, “now you would not hide behind these... at the same time pass over in silence his very large one” (http://oll.libertyfund.org/titles/galilei-...deserve to know the truth about our world. Jonathan Aguilar (@AggleDetector on Twitter) is a graduate student...

...and uses the similar buffers as would be used for a mini prep, except it does it on a larger scale. Using...for standard molecular biology techniques Browse our video library Check out our molecular biology reference...able to do high throughput DNA preps and other molecular work if we need it to. This gives us the oppotunity...the Biomek FxP is it’s customizability. It has a large deck that can house many different types of devices... DNA purification We're developing ways to prep large quantities of DNA because some countries just can...

... inherited by daughter cells during cellular reproduction, similar to the host’s chromosome, or they can...both Archaea and Eukaryota can also harbor these circular DNA passengers. Most natural plasmids encode beneficial...transfer can occur naturally through transformation (cellular uptake from the environment), transduction (viral... in a minute! Fun fact! Electroporation is one popular method scientists use to promote transformation...encoding these functions form the basis of many molecular biology tools! Figure 1: Common functions... Mother Nature’s toolbox for new and exciting molecular technologies!  This post was originally written...

...to be large enough to target larger areas, typically a few hundred nl. Alternatively, for larger brain...activity, commonly via light-activated ion channels. Similarly, chemogenetic tools are designed channels that...also reduce expression levels of certain genes. A popular method uses Cre recombinase on transgenic flox ...lentiviral vectors are superior packaging vectors for larger genes - they can hold up to 8 kb, while AAV can...liquor of the cerebral ventricles (ICV, intracerebroventricular) to minimize tissue damage; however, the...

...Biodesign Center for Molecular Design and Biomimetics and the School of Molecular Sciences at Arizona ...In response, the World Health Organization has declared a global health emergency and called for the fast-tracked...sensors within an extensive infrastructure of molecular and electronic companion technologies that has...1x106-fold lower than before), and our combined biomolecular tools can distinguish between global strains...detect the virus in a clinically relevant setting. Molecular diagnostics for the Zika virus predominantly look...Low-Cost Detection of Zika Virus Using Programmable Biomolecular Components." Cell (2016). PubMed PMID: 27160350...

...coding the molecular biology of the worm into a computer model that recapitulates how molecular biology ...get involved a in variety of aspects from the molecular biology to learning how to grow and care for the...but we need to develop the manual skills at the regular microscope, and the scripting skills at the automated...locomotion. Our most recent models of C. elegans molecular biology are based on the standard worm genome....Center (CGC) at the University of Minnesota has declared our makerspace an educational institution meaning...Silicon Valley as a student and then instructor at Singularity University at NASA Ames, and an instructor at...

...250 kDa Similar in size 8% 20–250 kDa Similar in size 10% 10–100 kDa Similar in size ...Conversely, if your protein of interest is large, look for a larger pore size (0.45 µm).  Types of transfers...important part of your western is your antibodies, particularly your primary antibodies. We always recommend...When choosing your lysis buffer, consider the subcellular location of your protein. Proteins located in...to overnight to run, and is recommended for very large proteins. A semi-dry transfer is faster — 3–30 minutes...differing amounts of available epitopes for that particular antibody, this truly does provide a relative,...

...noise, due to the large amount of dsDNA in most starting materials, and is not particularly sensitive. The...Protocol: How to Design a primer  Molecular Biology Reference Molecular Cloning Techniques References Bell... copy number assays for years. qPCR works like regular PCR, just with signaling molecules added to the...be affected by DNA quality and primer design, particularly for amplicons under 100 bp (Ma, 2015.) A schematic...ll have to consider:  Resources needed Most molecular biology labs and departments will have qPCR machines...

...columns had similar DNA yields. Additionally, columns regenerated up to five times had similar DNA yields...Addgene.org Find protocols for molecular biology and cloning Read our Molecular Biology Reference Visit our...plasmid miniprep protocol from Addgene follows a similar workflow as a column-based plasmid extraction kit... This method provides DNA yield and quality similar to that obtained with commercial kits, but in less...cellulose-based method was faster, cheaper, and had a similar level of sensitivity. Figure 2: Steps to ...

...fluorophores conjugated to molecular species with high affinity for endogenous molecular targets such as paclitaxel...conjugate, and the complexity of the system. A particularly desirable property for small-molecule labeling...removing excess dye from the sample, which is particularly important for samples where washing is difficult...and fluorogenic ligands. New fluorophores As a corollary to these innovative labeling strategies, several... these fluorophores are often bulky and contain polar groups (Figure 2b), which can preclude their use... and structure at the single-molecule level." Molecular cell 58.4 (2015): 644-659. PubMed PMID: 26000849...

...Molecular cloning requires some method of screening colonies for the presence of an insert. Traditionally...the insert site. A positive clone will produce a larger size product then a negative clone without the ...determine their size. Make sure to use an appropriate molecular weight standard for reference and to add a loading...-) will not. Backbone-specific primers (2) give larger sized products for positive clones (+) compared..., colony PCR is a useful tool to have in your molecular biology tool kit. Consider giving it a try next...Tips and tricks from the bench Don't pick too large of a colony. Too many bacteria can inhibit your ...Resources on Addgene.org Read about Choosing a Molecular Cloning Technique Learn to Perform a Bacterial...

... looking for some additional information or clarification on these vectors. Read on to find the answers...allow researchers to target transduction to a particular cell type or tissues; this is useful in gene ...therapy applications where a distinct subset of the cellular population needs to be targeted. Q6: Are transfer...self-inactivating, which is achieved by deleting a large portion of the 3'LTR in the transfer plasmid. The...not only the transgene but also that of adjacent cellular genes. Q7: Which genes are deleted or modified...through insertional mutagenesis. These risks are largely based upon the vector system used and the transgene...Library Resources on Addgene.org Addgene's most popular lentiviral plasmids Overview of Lentiviral Packaging...

...codons altogether.  So what does this mean for a molecular biologist who wants to express genes from one ...might want to optimize codons for expression in a particular organism. Why is codon usage important? Translation...tRNAs to build a peptide. Image from Mariana Ruiz Villarreal. By the late 1980s, scientists even created... vivo (Slimko and Lester, 2003).  Addgene has a large selection of codon optimized genes, so check out...information encoded in the primary sequence of a particular gene which could affect translation initiation...optimization in human therapeutics. Trends in Molecular Medicine 20:604–613  https://doi.org/10.1016/j.molmed...Resources on Addgene.org Check out Addgene's Molecular Biology Reference Read about genetic code expansion...

...toolbox. However, both of these proteins are quite large — comprising approximately 1000–1400 amino acids...challenge for efficient delivery into cells, particularly via viral vectors. While viral delivery is a... cell types, it has a limited capacity to carry large DNA cargos. As a result, the search for smaller,... Cas12a, uses just one to cleave both strands. Similar to how Cas proteins require a PAM (protospacer ...capacity, which makes it difficult to accommodate the large coding sequence of commonly used nucleases like ..., Nasti, R. A., Vollbrecht, M., Starker, C. G., Clark, M. D., & Voytas, D. F. (2019). Plant gene editing...an Old Favorite A History of Genome Editing in Popular Culture Additional Resources on addgene.org Genome...

...regions. PHP.eB, an enhanced version of PHP.B, shows similar transduction efficiency, but at a log lower dose...insertions and deletions (indels) most likely to have large phenotypic effects such as those that affected gene...pattern in the mouse brain.  Both labs then took similar routes to verify LY6A’s role as a receptor for ...transduce cells. The Wilson Lab generated results similar to the Deverman Lab when they looked at LY6A protein... B Transport across the Blood-Brain Barrier." Molecular Therapy 27.5 (2019): 912-921. PubMed PMID: 30819613... of AAV-PHP. B are limited to C57BL/6J mice." Molecular Therapy 26.3 (2018): 664-668. PubMed PMID: 29428298... across the blood-brain barrier: LY6A, a novel cellular receptor for AAV-PHP. B capsids." bioRxiv (2019...