We narrowed to 798 results for: REP

... materials in our repository. We started to increase accessibility to our repository by making it easier...Addgene opened in 2004, we focused on creating a repository that made it easy to share plasmids, both for...to — so far, we’ve shipped materials from our repository to an amazing 111 countries (Figure 1)! But while...that temperature-sensitive materials like viral preps and libraries were kept at the appropriate temperature...Systems AAV Plasmids Blogs Origin of Replication What is a Plasmid? How to Design Your gRNA for...Addgene blog in 2023. Views from a country are represented in blue; darker blue indicates more views from...YouTube channel, 2023. Views from a country are represented in blue; darker blue indicates more views from...

...the new plasmids, antibodies, and viral preps in the repository through our Hot Plasmids articles.  Here's...gene disruption in M. abscessus  Bioluminescent reporter of mechanical forces   Epitope tag antibody ...Western Blot for Anti-V5 [IPI-SV5-Pk1]. Addgene Report, https://doi.org/10.57733/addgene.l0ulu4. Hanke...through eight stages of embryonic development. Representative images of single focal planes through the middle...across species due to differences in natural DNA repair pathways and spontaneous mutations.  Here, the ...results in either bacterial death or indels after DNA repair. B) M. abscessus is first transformed with Cas9...an integrative plasmid containing tetracycline repressor TetR and resistance selection marker KanR. This...

...resistance gene and the origin of replication are necessary to replicate plasmids and make sure the cells... operon Inducible by arabinose and repressed catabolite repression in the presence of glucose or by competitive...transcribed promoters Inducible with galactose; repressible with glucose  Can be used independently or together...mRNA Yeast promoter for alcohol dehydrogenase I Repressed by ethanol Full length version is strong with ...expression Promoter from E. coli tryptophan operon Repressible Gets turned off with high levels of cellular ...lac operon Constitutive in the absense of lac repressor (lacI or lacIq). Can be induced by IPTG or lactose...expression. lacIq mutation increases expression of the repressor 10x, thus tightening regulation of lac promoter...

...Democrats more often than Republicans, the March is not about Democrats or Republicans (16). This is a Pro-...contact your representatives in case you would like to talk with them about science: Representatives: To find...contact information for your representative in the House of Representatives all you need to know is your...in this article are those of the author do not represent a formal stance taken by Addgene or its staff....Climate change Vaccines GMOs Transgender issues Reproductive health Immigration Pollution Healthcare Science...Legislatures: To find the website to contact representatives at the state level you can click on your state...

...ready-to-use viral vector preps. Today, that number has grown to over 114,600 viral preps. And in 2025, we’ll...service would allow people to add a gene to a repository others could request from — Addgene!  Addgene...meant we could grow accessibility alongside our repository.  “I’m sure you hear it all the time, but Addgene...took almost six years to reach our first major repository milestone — 10,000 plasmids deposited and available...conferences. And when CRISPR arrived in 2012, our repository was rapidly able to support sharing of CRISPR...milestone! We were thrilled to see the impact our repository was having on the scientific community, but it...-quality service, open data, and increased reproducibility of materials. In March 2022, we launched our...

...random variation, or noise, in reporter expression. To do so, one places reporter expression under the control...expression of a reporter protein, it is possible to change the amount of noise in reporter gene expression...highlight a subset of the new plasmids in the repository through our hot plasmids articles. These articles...deliver the target antibody. TRIM-Away reagents and reporters are available from Addgene - we hope they will... been fused to a variety of protein modules to repress or activate gene expression, sometimes incorporating...lab has deposited plasmids encoding fluorescent reporters and other proteins controlled by GAL4 and ZIF268...progesterone controlled transcription factor as well as reporters specific to each transcription factor. They then...

...and utilization - and on self-propagation by reproduction. Both physiological processes are controlled...ligands. Nuclear receptors and their ligands NRs represent the largest family of transcription factors found...expression of genes involved in a broad range of reproductive, developmental, metabolic, and immune response...more NRs for efficient and timely activation or repression of gene expression (see www.nursa.org for a curated...Coregulators can function as coactivators and corepressors depending upon a context defined by ligand, ... third phase of funding, the NURSA Consortium represents a unique funding model in nuclear receptor research...the development of a vast number expression and reporter plasmids and the free exchange of these vectors...

...additional origins of replication compared to monomers, they are also able to replicate at a higher frequency...restriction sites in multimers usually repeat in tandem along with the repeated plasmid sequence, giving greater...much longer than expected, and the sequence was repeated two or more times in tandem. Such plasmids are...produces a multimeric form of the plasmid with repeating sequences. In some specific cases, multimerization...large inserts (>7 kb)(Berg et al., 1989), direct repeats (Ribeiro et al., 2009), and at high copy numbers...<300 bp) which cannot distinguish between large repeat sequences within a plasmid, it cannot be used to...sequencing. These longer reads are able to detect large repeat sequences within a plasmid and, therefore, allow...

...process ensures that any potential sequence discrepancies can be caught prior to distribution. Catching...issue? At Addgene, a QC issue is defined as a discrepancy that we believe may affect the function of the...decisions when analyzing the wide variety of discrepancies encountered in plasmids deposited to Addgene...resolve. Sequencing and assembly issues Sequence discrepancies are not always a result of mutations in the ... platform, such as GC-rich regions and highly repetitive sequences and/or features. Most of these regions...will contact the depositing lab to review the discrepancies. Often, the identified issues will not affect...requesting scientists aware of the identified discrepancy in the plasmid, and that it is not known to impact...

...the new plasmids, antibodies, and viral preps in the repository through our Hot Plasmids articles.  Here's...PAGE editing with viral sgRNA targeting mCherry reporter (mChe), with co-incubation with assist peptide...variant (Piatkevich et al., 2023). Check out their preprint for more details. Not only will these mStayGold...doi.org/10.1038/s41592-023-02085-6. PMID: 38036853. Preprint: https://doi.org/10.21203/rs.3.rs-2941917/v1.  ...doi.org/10.1038/s41587-023-02018-w. PMID: 38081970. Preprint: https://doi.org/10.21203/rs.3.rs-2684100/v1.  ...proteins derived from StayGold. ResearchSquare (Preprint). https://doi.org/10.21203/rs.3.rs-3188559/v1..... CAP-Mac also robustly expressed fluorescent reporters in green monkeys and cultured human neurons, with...

...sure that what we observe through the microscope represents reality as accurately as possible. This is especially...we have created an initial set of 10 plasmids representing 10 human genes and will add more to the collection...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 cells, ...majority of the time the genome will attempt to repair itself without outside influence (non-homologous... the stem cell will seek out a template for the repair process. By flooding the cell with a plasmid containing...cell into using the introduced donor plasmid as a repair template. This results in the insertion of the ...

..., E4, and VA RNA genes; and the pRepCap plasmid containing replication factors and serotype-specific capsid...lentivirus, AAV does not possess the ability to replicate itself and depends on helper viruses like adenovirus...herpes simplex virus to facilitate efficient replication. The most common production system, however, ...been developed to determine the titers of rAAV preparations, such as dot blot, enzyme-linked immunosorbent...different titers are measured. A TCID50 measurement reports an infectious titer, meaning the concentration ...targeting the viral genome or inverted terminal repeats (ITR). You should always have a high-quality plasmid...overestimate of transducible units in the virus preparation.  rAAVs  Now your AAV virions are ready to store...

...along with a reporter, such as a fluorescence gene or a resistance gene. These reporters allow you to ...multicistronic element(s), or you could replace one of the reporter genes above with your gene or genes of... using IRES elements. Further, scientists have reported lower expression of the downstream cistron due...eukaryotic cells, and, although some scientists report close to 100% cleavage with some of these peptides...that already has the multicistronic element and reporter cloned in. In these plasmids you would simply ... 2A element (depending on the placement of the reporter gene). Addgene's collection offers a variety of... are designed to have one or more of the genes replaced.  Additionally, 2A peptides could be PCR-cloned...

...that can be repaired by either non-homologous end joining (NHEJ) or homology directed repair (HDR). This...photoactivatable repressor (dCas9). The activity of all variants was reversible and repeatable. Since these... spatiotemporal control, reversibility, and repeatability. The system developed by the Sata lab is composed...could be spatially controlled in a reversible and repeatable manner. With their light-activated CRISPR/Cas9...The system was also shown to be reversible and repeatable when blue light (450nm) was cycled on-off-on....system, Ye’s lab could edit a tdTomato fluorescent reporter gene in mice using Minicircle DNA vectors encoding...seen in the figure below. Next, by using a dual reporter fluorescence assay, Hemphill et al. demonstrated...

... countries, into our repository, while expanding global access to our repository through our distributor...At 3500 plasmids, this represents the largest single deposit in our repository and was a three–year project...14 deposits, that became available through our repository in 2023.  In June, we announced a new partnership... launch our Antibody Data Hub, an open-access repository for antibody validation data. We’ll continue ...expanding international access to the Addgene repository, partially through work with our distributors...we are now able to ship rabies plasmids in our repository to international requestors.    I would be remiss... used by our requestors, and supported by our repository. You can enjoy these stories, and other 20th ...

...Sanger sequencing to confirm an insert that is repetitive or otherwise hard to sequence using NGS or long-read...such as G-C rich areas, secondary structures, or repetitive regions. Short-read sequencing can also be time-consuming...time-consuming, depending on the library prep and instrumentation used.  Here at Addgene, we sequence ...sequencing or Sanger sequencing, either because of repetitive regions or for higher accuracy of a particular...This method is great for resolving complex and repetitive sequences, but can be costly, low-throughput,...Nanopore sequencing to QC plasmids that are highly repetitive, such as those containing cassettes of the same...the reads, which boosts ‌accuracy, especially in repeated regions. If this doesn’t resolve the assembly ...

...storage, viral preps should be divided into single-use aliquots, and stored at -80°C. Some reports suggest rapidly... time consuming, difficult, and lacking in reproducibility. Read on for some tips for getting the most...Obtain a vial of the desired cell line from a reputable distributor such as the American Type Culture ...you may want to consider concentrating your viral prep. Storage can also play a role in the success of ..., it is best to avoid freeze-thawing lentiviral preps unless absolutely necessary. Many labs try to use...this blog post for tips on titering your lentiviral prep!) and infect. While this approach is a great start...transducing with a range of dilutions of your lentiviral prep. Hopefully, by using more concentrated dilutions...

...sites if in double stranded DNA are repaired by base excision repair, but can lead to mutations in single...noticed that there are many options to elute your DNA prep. You might see protocols that recommend eluting ...single stranded DNA undergoing replication.   DNase on the other hand can digest and degrade the double ... buffer for preserving the stability of your preparation for a long time. Tris buffer controls the pH,... if the priority is to directly use your DNA preparation in any downstream application involving enzymatic...buffer is that it allows better control of the pH preparation stabilizing the plasmid, and still allowing most...at -20°C or even at -80°C, in this case your preparation can be eluted in water or in your buffer of preference...

...confirm the identity and serotype of our AAV preparations. We recently published a description of the ... For labs that produce a small number of AAV preparations this level of quality control is sufficient ...facilities producing a large number of viral preparations in parallel, there is a very real risk of mixing...AAVs and the TA-based ligation and tagmentation preparation approaches described previously require a preliminary...effective substrates for tagmentation-based library preparation procedures. This finding eliminates the double-stranding...simplified approach, we sequenced over 100 AAV vector preparations and showed that VGS can quickly and reliably... script that can confirm the serotype of AAV preparations.  Using viral genome sequencing to determine...

...platform for scientists using Addgene AAV viral preps to easily find and share useful technical data. ...tools. This word cloud (using wordart.com) represents contributed data on brain injection site from...15 minutes or less if you have all of your data prepared. All of these questions relate to factors that...that you should have already recorded as good reproducible and record-keeping practice. In fact, when you...did you use? For animal subjects it is helpful to report the genetic background and catalog number or RRID...exactly which strain you used so if they wish to replicate your experiment, they know exactly where to get...take negative data, which is equally important to report. We recently published a paper in Frontiers in ...