We narrowed to 1,052 results for: fer

...you can quickly test that part in different contexts and plug different characterized parts into a TU! What...works on genetic code expansion.  References and Resources References Weber E, Engler C, Gruetzner R, Werner...cloning is MoClo. That’s because MoClo specifically refers to the organizational schema described above, and...nature of MoClo allows users to test a variety of different orientations and components within the system ... a great tool to deliver many combinations of different candidate genes at once to identify the best cocktail...and gene editing that use MoClo, for which many different MoClo kits have already been established. Keep...

...Figure 9: epegRNAs (left) and PE7 (right) offer two different strategies for protecting pegRNAs from degradation...software in the comments below! References and Resources References Anzalone, A. V., Randolph, P. B.,... These tips are a great starting point: Test different lengths of the primer binding site, starting with...still be optimized (Anzalone et al., 2019). Test different lengths of reverse transcriptase template, starting... templates, it is even more important to test different lengths and sequences, as unintended secondary...specific prime editing tools There are a lot of different prime editing tools to choose from, and many have...

...tools for genetic engineering. There are several different viral vectors to choose from, for example gamma-retrovirus...separate envelope and packaging plasmids as well as transfer plasmids. This concept ensures safe handling and...isoforms of these proteins, as well as the viral LTRs differ. Therefore, lentiviral and gamma-retroviral packaging...Gamma-retroviral vectors Popular gamma-Retroviral Transfer Vectors   Expressing your gene of interest pMKO...expressing RNAi pMKO.1 puro (Plasmid #8452) RNA interference (RNAi) is an RNA-mediated gene silencing mechanism...interest. Lentiviral vectors Popular Lentiviral Transfer Vectors    Expressing your gene of interest pLenti-puro... in phenotype. The plasmid comes with a 1.9 kb stuffer sequence that can be removed and replaced with ...

...pathological conditions. References: Monticelli, M., Ferro, T., Jaeken, J., dos Reis Ferreira, V. & Videira, P...GdnHCl to patients suffering from rare diseases arising from Arg mutations. References: Baldwin, Enoch,...life-threatening consequences. They result from defects in different steps in the glycan biosynthetic pathway and have...recently reviewed and reported that PMM2-CDG patients suffer from immunological defects. These are characterized... discovered that the in vitro functions of 11 different Arg-mutant enzymes could be rescued by supplementation...

...collaborate with specialists in different fields that can provide different insight into an area - not just...you are looking to move to a different university, perhaps in a different country, choose where you go...collaborators It’s a good idea to move around to different labs during your career; you learn new techniques...I hope to discover. This section may make the difference in whether you get funding or not; applications... grants too There are actually quite a lot of different pots of money available for researchers, so look...gained in writing small travel grants to attend conferences and a seed funding grant for a proof-of-principal...

...publish a new blog post, and live tweet from conferences. Find more tips on using social media as a scientist...for example at an anniversary or for Girls’ Day, offer guided tours to show people where the lab magic ...communicate science. You can draw live sketches at conferences like @ATJCagan or create agar art with colorful...Association, EUSJA or ABSW for helpful tips and job offers. If you live in the UK, join the SciComm mailing...10 days. And the Society of Experimental Biology offers press internships to cover their annual meetings...College London. Similarly, the University of Cardiff offers an MSc in Science, Media and Communication. In ...Santa Cruz, MIT, UW Madison, and Boston University offer science communication programs. Others, like Northwestern...

... can only carry around 4.5 kb. Different AAV serotypes have preferences for specific cell types. For example...behavioral studies. References Stoica L, Ahmed SS, Gao G, Sena-Esteves M. Gene transfer to the CNS using ...the experiment. Handling can however negatively interfere with certain tests and is not always recommended...end of an experiment series would behave very differently (distracted) from the ones tested at the beginning...Experiment Behavioral scoring systems (experimenter differences in behavior observation); experimenter blinding...injections. AAV particles are smaller and may be preferable if wider diffusion is required. On the other ...

...these will not be detailed here, but additional references are provided at the end.  Nearly all common bacterial..., and III), each responsible for transcribing different types of RNA: Polymerase I is responsible for ...that each eukaryotic RNA polymerase terminates differently. Polymerase III, for example, relies on a specific...Rho-independent termination found in prokaryotes. This is different than Polymerases I and II, which both require ...factor dependent, Polymerases I and II employ different mechanisms to terminate transcription. Polymerase...and/or use weaker poly(A) signals like BGH.  References Peters JM, Vangeloff AD, Landick R. Bacterial...Promoter-Dependent Manner. Hum Gene Ther. 2008. PMID: 18627247 Zufferey R, Donello JE, Trono D, Hope TJ. Woodchuck Hepatitis...

... optical insulator called a ferrule (below). The fiber optic cable/ferrule is inserted into the target...optic patch cable is then connected from laser to ferrule to deliver light pulses to the target brain region...Several protocols have been written outlining the different steps and materials required for each [1, 2]. ...photons from the laser to the patch cable to the ferrule implant must be maintained during the entire protocol...motor cortex of a mouse. Upon connection of the ferrule implant to the laser with the patch cable (difficult...discussed above, each opsin variant has slightly different activation/inactivation kinetics. For example,... or follow him on Twitter @derekpsimonphd.    References 1. Gutierrez DV, et al. Optogenetic control of...

...advantages SapTrap offers for tagging a single genetic locus, the toolkit also offers repair templates ...improve CRISPR genomic tagging in C. elegans. SapTrap offers scientists a convenient one tube assembly reaction...will produce the same unique SapI 5’ overhang, different combinations of connector and tag donor plasmids...donor plasmids for protein tagging, including luciferase (NanoLuc), fluorescent proteins (TagGFP2, TagBFP...cell line stably expressing Cas9. This PCR toolkit offers C- and N-terminal tagging vectors (eg GFP, Flag...protein expression levels. CRISPR tagging systems offer users an easy and efficient way to tag endogenous...light on previously unknown protein functions. References Schwartz, Matthew L. and Eric M. Jorgensen. “...

...yours does and then use that time! Different platforms have different quirks, so remember to practice on...that requires its students to do rotations in different labs before selecting an advisor, it is always...! Prepping for the interview Many universities offer free practice sessions for students doing virtual...points and keeping them at-hand, you can quickly reference your questions about the PI’s research or the ...the name of the award you won for your conference presentation. It can help soothe your nerves and make ...like sickness or scheduling issues can and do interfere with those in-person plans. Virtual options are...good fit for you. The application process looks different than it did a few years ago, but I hope the tips...

...extraordinary molecular detail.   References 1.Patterson, George H., and Jennifer Lippincott-Schwartz. "A photoactivatable...Fluorescent proteins (FPs) offer scientists a simple yet powerful way to tag cellular proteins and investigate...most scientists working on in vivo imaging models perfer the PA-RFPs over PA-GFP. Another class of irreversible...dimeric or tandem dimer versions of EosFP are preferred over the monomeric one since the formation of ...organelle trafficking. Monomeric PA-FPs are usually preferred for studying single protein characteristics, as... as the monomeric state is less likely to interfere with endogenous protein structure and function. By...non-invasive labelling, but in contrast, PA-FPs offer a reduced potential for photobleaching and phototoxicity...

...helping everybody, helps everybody! References and resources References Dayton, R. D., Grames, M. S., & ...scientists under-informed with respect to how different serotypes, promoters, and injection methods are...published.    Fig. 1: Testing of three different AAV serotypes (AAV1, 8, 9) injected into the A2...biological reagents; many factors, like storage buffers and conditions (3,5,8), injection titers (1,2),...literature, scientists can use the Data Hub to compare different methods and vectors, and use this data to guide...& Klein, R. L. (2018). More expansive gene transfer to the rat CNS: AAV PHP.EB vector dose–response and...sharing practices and data availability upon request differ across scientific disciplines. Scientific Data,...

...most cases, interested in transferring genes to mammalian cells (i.e. transferring DNA), yet retroviruses...lentiviral vectors. Lentiviruses are sometimes referred to as “complex retroviruses ” (Cullen, 1992) because...generalize the idea of “transduction” to mean any transfer of material from the virus to the host. In fact...pseudotransduction” — to describe the direct protein transfer of RT, which would actually occur in step 2 during...dimerized linear ssRNA strands (which can have a different genomic sequence from one another) — by various... with lentiviral replication.  Resources and References More resources on the Addgene blog Viral Vectors...popular lentiviral plasmids γ-Retrovirus Guide References Cullen, B. R. (1992). Mechanism of action of ...

...to the plasmid search results: More than 200 different GCaMP containing plasmids are available for distribution...biosensor using AAV, you might decide you’d really prefer to order ready-to-use virus rather than producing...starting a new search or clicking through to a different part of the website, you can now use the pulldown...pulldown menu on the search page to quickly jump to different types of items: Did you notice the “Filter By.... The following screenshots show the range of different terms we can use to find materials from this publication...dropdown menus below the search bar. As you try different terms you may find the available categories and...you get this far and realize you need to apply different filters you do not need to start a new search....

... number of different strains of E. coli that can be used for transformation. The preferred E. coli strains...freezers that the samples can be stored in? Different shelves? Different racks on each shelf? Give each container...long-term storage in our facilities, every sample is transferred to a 2D barcoded tube (see next section) and ...about location, location, location There are many different types of samples at Addgene - DNA sent from depositors... concentration or need to be transformed into different cells than were originally requested. Most of ...analysis from our protocols and molecular biology reference pages! Resources Avaliable on the Addgene Blog...

... protein as pure as possible? Vectors that offer different combinations of tags are readily available,...peptide-based proteosome inhibitors, glutathione S-transferase (GST), which can be fused to recombinant proteins...Purification Binding and elution steps use very moderate buffer conditions FLAG DYKDDDD or DYKDDDDK or DYKDDDK...co-IP, IF, flow -cytometry; can occassionally interfere with protein folding HBH HHHHHHAGKA GEGEIPAPLA...small size, rarely affects function S-tag KETAAAKFERQHMDS  1.8 Solubility and Affinity Abundance of charged...common. Tandem affinity purification (TAP) once referred specifically to a combo tag comprised of a calmodulin...information and some protocols can be found in the references provided. Additional resources: Young CL, Britton...

...delivered on a different plasmid and allows simultaneous multiplex editing of six different genes. This ... Jan 28, 2016 and last updated Sep 10, 2020 by Jennifer Tsang. CRISPR makes it easy to target multiple...flanking the promoter-gRNA construct, but with different sequences adjacent to the sites. When digested...assembly principles, but they’re optimized for different purposes.   Gersbach Lab multiplexing plasmids...kanamycin-resistant plasmids, each containing a different gRNA target sequence downstream of the 7SK, human...users to express up to 7 gRNAs. The kit contains different destination vectors depending on the total number...transcript. Rather than being transcribed from different promoters, the gRNAs are transcribed together ...

...between Cas9 enzymes from different species, explaining why they recognize different guide RNAs. Deep in the...Other CRISPR enzymes like Cas12 have different structures and different uses. In all cases, structural biology...techniques, like electron microscopy and FRET, can offer more information on these regions.   Cas9’s transition...from the same angle to highlight conformational differences. HNH, red; RuvC, orange; CTD, yellow; REC lobe...research. New details are still being uncovered, and different techniques all bring their own advantages—and ...Allergy and Infectious Diseases. Resources and references More resources on addgene.org  Addgene's CRISPR...656. https://doi.org/10.1038/s41579-022-00739-4 References Bravo, J. P. K., Liu, M.-S., Hibshman, G. N.,...

...Vector Core. Transfections are performed using the transfer plasmid, a plasmid encoding rep and serotype-specific...taken offline and discarded. Confirmation of the transfer plasmid Addgene uses a rigorous barcode matching...preparation undergoes PCR with primers targeting the transfer plasmid used in the transfection. PCR products... matching system and PCR-based confirmation of transfer plasmids used in Addgene’s viral production. Barcoding...After production, we confirm the identity of the transfer plasmid in the viral preparation by performing...performing PCR against unique regions of the transfer plasmid.   Lentivirus resources Browse lentiviral preps... by Ina Ersing in August 2023. Resources and references Resources at the Addgene blog Find and share ...