We narrowed to 1,558 results for: TRI

... by ultracentrifugation (75,000 - 225,000 g for 1.5 – 4 hours at 4 °C). After ultracentrifugation, virus...integrity of your virus, perform a diagnostic restriction enzyme digest and compare it to some reliable...vectors) you can concentrate your virus by ultracentrifugation of the viral stock, followed by resuspension...packaging cell debris before proceeding to ultracentrifugation to avoid contaminating your transduction ...through a 0.45 µm or 0.22 µm filter) or by a centrifugation step (5 minutes at 300-500 g). After removing... to concentrate a virus solution without ultracentrifugation, you can reduce the volume of cell culture...

...days Purification Ultracentrifugation (AAV); centrifugation (lenti) Centrifugation Titration time ~...using methods like ultracentrifugation, while lentivirus supernatants are centrifuged and filtered to eliminate...have a higher titer than a P1 virus. Pro-tip! Centrifuge the viral supernatant in between passages to ...straightforward — simply spin down the supernatant in a centrifuge to remove any unwanted cellular debris, as you...

...and sensing electrical potential. Thankfully, there is a tool to measure these electrical events! Voltage...biological tools. Why Use Voltage Indicators? Electrical signals are integral to many facets of biology...muscle contraction – is directly related to the electrical impulse of the cardiac cells. To study this phenomenon...is carried out by action potentials which are electrical charges propagated through multiple cells to ...depends on the ability to monitor their signals – electricity! The research tools for studying voltage within...was dipicrylamine, a lipophilic compound that distributes itself within the plasma membrane as well. Another...

...accelerate your research.   Why distribute virus? Plasmid distribution is booming, but, over the years... we can help them share and distribute other reagents. Virus distribution came up time and time again ...Addgene! As part of our new Viral Service, we’re distributing lentivirus (with many CRISPR tools included ...with light will be available soon. We’re also distributing AAV preps with particular serotypes that can...here. If you have virus expertise you’d like to contribute to the blog, please email us and we’ll get you...quality control data for each of the materials we distribute. Quality control info can be found on each viral... the Viral Service or any of the materials we distribute, please send an email to [email protected]. Your...

...likely to be positively impacted by the trial. During clinical trials, expansion of participants based on ...This post was contributed by guest blogger Subhadra Jayaraman, a doctoral candidate at Binghamton University...surveillance. Dormancy of certain cancers can be attributed to the concept of immunoediting, by way of favorably...TGFβ. T-regs are suppressive immune cells that restrict the production and activity of other effector ...endothelial growth factors (VEGF) in tumors is attributed to a signaling cascade, where lack of oxygen ...therapies include radiation therapy (owing to contributions from Madame Curie), chemotherapy, and surgical...leukemia and lymphoma and are in their clinical trial phase. In all of the above therapeutics, techniques...

...encouraging user contributions (using simple web forms) while also maintaining a relatively strict database design...This post was contributed by guest blogger Talley Lambert, a Research Associate at Harvard Medical School...design of an imaging experiment.  The choice is non-trivial, however, as FPs are tremendously complicated entities...on a web-based interactive visualization of FP attributes (which turned into www.fpvis.org).  That page...and we very much appreciate even those small contributions). Of course, no community-editable database ...simplifies the process of building advanced multi-parametric search queries.  Looking for a bright, monomeric...frequently) for the sake of comparison or ease of retrieval.  These collection pages can be kept private, ...

...their destination co- or post-translationally. TRICK: Visualizing the first round of translation While... co-opts the physical location of the ribosome, TRICK (Translating RNA Imaging by Coat protein Knock-off...RNA-binding proteins to allow translation to occur. For a TRICK reporter mRNA this displacement leads to a change...RNAs appear red. To enable this color switching, TRICK requires the following components: a GFP protein...translation is necessary for removal of GFP from the TRICK reporter. Delayed translation is known to be important...important in development, and Halstead et al. used TRICK to examine translation of Drosophila patterning ...patterning gene oskar. Early in development, the osk-TRICK reporter was double labeled, indicating translational...

...This post was contributed by Jake Watson and Javier García-Nafría from the MRC Laboratory of Molecular...present even in common lab cloning strains. This intrinsic bacterial recombination pathway, referred to as... longer oligos, which could be avoided using restriction enzyme cloning, were more prohibitive at the ... doesn’t matter if the DNA is PCR amplified, restriction digested or even a synthesized gene. PCR is the... Mutagenesis is performed by designing primers astride the old codon, amplifying the whole vector. The...extending homologous sequences (to 25-30 bp). Tips and tricks for IVA cloning Use a sequence manager A sequence...IVA can be employed by cutting the vector with restriction enzymes and adding the corresponding homologous...

...colonies in petri dishes, but close to 80% of samples are received as DNA in a microcentrifuge tube. Once...sample enters Addgene’s lab, a series of events is triggered. Each step is tracked through barcoded tubes that...track all of our 40,000 samples available for distribution!    Check Out How We Think about Developing...The preferred E. coli strains for storage and distribution at Addgene are DH5α, NEB stable, ccdb survival...Each sample that is transformed is plated onto a petri dish plate that is also barcoded. The barcodes on... to be transformed with a micropulser using an electric charge to help the DNA enter the cells. Some samples...

...fragments, but uses site specific restriction sites (Type IIS restriction endonucleases) to cleave DNA outside...which have provided appealing alternatives to restriction enzyme cloning. These newer technologies have...They offer many advantages over the traditional restriction enzyme cloning we once relied exclusively on....that does not require the presence of specific restriction sites (or lack thereof) nor a serious time commitment.... If you inserted these two pieces of DNA by restriction enzyme cloning, you would have to do it in two...the smaller overlapping sequences left with a restriction digest. A study in 2013 found that Gibson assembly...PCR step difficult--OR there is no convenient restriction site for linearization. This is a perfect case...

...otherwise requested. Distribute pending QC: your plasmids become available for distribution as soon as they...archive any plasmids you've deposited with us and distribute them to scientists worldwide. What's more, depositing... pre-publication plasmids" and set a desired distribution status. You can submit "Published plasmids":...description of the deposit. Select your desired distribution status: Hold for publication: we will initiate... us with publication details or permission to distribute, or Addgene finds your journal publication describing...additional questions. Bacterial Strain Addgene can distribute your bacterial strains if they: are E. coli ... +1 (617)-ADDGENE (233-4363) if you’d like to distribute your bacterial strain(s) through Addgene. Description...

...This post was contributed by guest blogger Derek Simon. The actual experiments you do will be determined...implanting an electrode and non-specifically electrically stimulating an ill-defined region of the brain...trained to “freeze” upon anticipation of a mild electric foot shock that has become associated with a visual...using HR was found to block the acquisition and retrieval of contextual fear memories [5].  In a modified...other regions of the brain may be involved in retrieval of remote contextual memories. This study is an...real-time inhibition), and behavioral (remote memory retrieval) specificity. Optogenetics can be used to more...2012): 159-172.5. Goshen I, et al. Dynamics of retrieval strategies for remote memories. Cell. 2011;147...

...that plasmid deposits come from 49 countries* and reached 102 countries. We started from 474 plasmid deposits...deposits coming from two new countries bringing the total to 51 countries with plasmid deposits!  ...Twitter!  That’s today through Friday. There will be trivia, prizes, and more! We wanted to take a week to ...impact in open science. Depositors consistently contribute to our collection so that we can offer the scientific...been requested about 1,500 times from labs in 45 countries*. While the bulk of these plasmid deposits came...you are a depositor or not. We will have Addgene trivia (with prizes!), polls, and shout outs, and we’ll...the schedule for the week: Monday: Fun stats and trivia begins! Tuesday: Depositor features (plasmid deposits...

...Optogenetic control of CRISPR-Cas9 by CASANOVA Article contributed by Cary Valley  Listen to the anti-CRISPR podcast...fluorescent voltage indicator Archon Article contributed by Shreya Vedantam  Listen to the voltage indicator...segment The neurons in the brain compute using electrical impulses to coordinate emotions, thoughts, and... behavior. Historically, scientists study the electrical activity in the brain by inserting electrodes...thousands of proteins for one that can report electrical activity via imaging. They took the light-sensitive...easier with a 2in1 vector cloning system Article contributed by Michelle Cronin  Listen to the 2in1 cloning...pUC57-L3L2) that offer the advantage of using restriction digest cloning to introduce the GOIs into the...

...anode/iblot gel matrix, the blotting membrane, the gel, a copper cathode/iblot gel matrix, and filter paper...remain intact and the protein is separated through a matrix by charge. In a denaturing western, the protein...smaller molecules moving more quickly through the matrix. For the purpose of this blog, we will be focusing...confirm the size of the protein of interest. An electric current applied to the gel causes the negatively...proteins encounter less resistance in the acrylamide matrix and migrate faster through the gel. For better ... In addition, PVDF membranes can be repeatedly stripped of an antibody and reprobed with different antibodies...sandwiched” together with blotting paper and an electric current is applied causing proteins to migrate...

...fundamental steps: creation of the RNAi trigger delivering the RNAi trigger characterizing the RNAi phenotype...This post was contributed by guest blogger, Robert Orr, who recently received a Ph.D. in Biology and ...complementary to your gene or genes of interest to trigger RNAi activation. A typical RNAi experiment can ...RNAi-based investigation. Creation of the RNAi trigger requires a few initial considerations, such as ...artificial microRNA). Using a DNA template for the RNAi trigger is the most prevalent method due to ease of use...assortment of siRNAs generated from a long hpRNA trigger, only a single ~21-nt interfering RNA is produced...precursor mentioned above and is a popular RNAi trigger in the mammalian research community. Fortunately...

...antibody (N206A/8 TC Supernatant). Image attribution: James Trimmer, UC Davis.   ...interact with the air-water interface (AWI) before vitrification, they can be completely denaturated (left) or...representative micrograph (C), particle orientation distribution and Fourier-shell correlation curve (D), and...protein. While the addition of LEAs may bias the distribution of particle orientations, this could be overcome...off-target editing. Cargo size limits are also less stringent for eVLPs than other viral delivery methods, allowing...anti-GFAP [N206A/8R] antibody shared by James Trimmer’s lab (Figure 5) as well as antibodies for identifying...

...study a matrix metalloproteinase: López-Pelegrín, Mar, et al. "A novel mechanism of latency in matrix metalloproteinases...Addgene. What you might not know is that Addgene distributes curated collections of plasmids as kits with ...This kit is designed to be used with Type IIS restriction enzymes and standardized primers to clone your... pExpression vectors) plasmids. The type IIS restriction sites used in these vectors are of low abundance...Biosafety of biotechnologically important microalgae: intrinsic suicide switch implementation in cyanobacterium...used to generate these fusions all use the same restriction enzymes for cloning. Furthermore, some plasmids...

...collaboration. Together, we distributed 184,824 materials to 39,529 scientists across 86 countries. Even more remarkably...remarkably, 1,160 labs from 522 organizations in 41 countries enriched our repository with 15,020 new plasmids...part of the story. This year, we made significant strides in making science more accessible and reliable....thoroughly characterized antibodies, and our contributions to key research papers are already inspiring...Errington, T. M., Turn, R. E., Bandrowski, A., Trimmer, J. S., Rego, M., Freedman, L. P., Ferrara, F.,...

...of months. Read on to discover what new post contributed the most to this record breaking month and to... Apps for Lab Life This very useful post was contributed by guest blogger Sean Stacey from Eastern Tennessee...Phage Genomes for Host Binding Proteins," was contributed by guest blogger Jessica Sacher from the University...Disease, Running for FOP, Running for AJ,” was contributed by Kurt Swanson, husband of Addgene Director ...currently on my way back from a very successful Outreach Trip in San Francisco and would like to give my thanks...who spoke with me and deposited plasmids on this trip. Here’s to hoping that October is even more awesome...