We narrowed to 645 results for: Rest

...selected the optimal promoter for your gene of interest, painstakingly cloned into the perfect empty backbone...DNA, important when trying to cut with certain restriction enzymes (ex: ClaI or XbaI) dcm DNA cytosine...DNA, important when trying to cut with certain restriction enzymes that are methylation sensitive. dnaJ...plasmid maintenance, blue/white screening, partly restriction-deficient; good strain for cloning repetitive...methylated, allows for methylation sensitive restriction enzymes to cut the plasmid after preparation....

...cloning sites for easy cloning of your genes of interest. In order to improve the study of specific neuronal...new AAV approach using regulatory elements that restrict expression to this cell type. To this end, they... ATP1A1 in addition to a researcher’s locus of interest. NHEJ- or HDR-induced mutations in ATP1A1 can ...locus increases the probability that the locus of interest has also been edited! Thus, ouabain selection ...the size of DNA fragments. Depending on which restriction enzyme is used to cut the plasmids encoding the...are shared through Addgene without licensing restrictions. Use these instructions to prepare the PSU ...

...pediatric kidney cancer. This was particularly interesting as I had just completed my thesis on the study... taking on a similar position in your field of interest before you dive straight into a full-time position...for bacterial contamination. It was a really interesting job at first, everything was very new, and everything...to help cover the costs. My MSc courses were interesting and luckily genetics is a sector which has been...the company as well and use my other skills and interests. I was so glad when my future supervisors were...

...developed the plasmids, pPSU1 and pPSU2. When restriction digested with PstI or EcoRV, these plasmids generate...significantly cheaper than commercial ladders. Are you interested in making your own DNA ladder? You can find the...available from Addgene. Learn How to Analyze a Restriction Digest References 1. Henrici, R. C., Pecen, ... Miniprep Protocol Learn How to Analyze Your Restriction Digests Make Your Agarose Gel Other Resources...

...dabbles in science policy, public outreach, and restorative justice.  Follow her on twitter @sung_vanessa...age live in a state with at least two abortion restrictions that conflict with scientific evidence. We need...Campus.  He studies viruses including HIV and is interested in global health policy.  He is also chair of...Michigan, where programs like the Great Lakes Restoration Initiative are at risk of major cuts and where...science and positively reinforce their scientific interests. Our marches across the world have had an impact...

...parts in a specific orientation using Type IIS restriction enzymes (Weber et al., 2011). It can be performed...units (TUs) used to express your favorite gene-of-interest, all the way to the complex multi-TU constructs...and tags makes EcoFlex a great option for those interested in recombinant protein purification and metabolic...looking to express high levels of their gene of interest in yeast should also take a look at the Yeast ...stacking, increasing production of a protein of interest, and improving the nutrient profiles within given...

...wanted to measure expression of your protein of interest in a single cell? Or perhaps, you need to analyze...fluidics system they pass through a flow cell which restricts the size of the stream and forces the cells to...? However, fusion proteins tag your protein of interest with a fluorescent reporter on the N- or C-terminus... around the specific population that they are interested in and further analyze that subset of cells. .... An immunologist, for example, might be interested in the lymphocytes and “gate” that population for ...

...always been interested in the ways that I could change the world around me. My strong interest in the intersections... marketing internship at Addgene. Although my interests are very clear, I had no idea how to translate...would be a perfect way of exploring my different interests but also a way of challenging what I am used to...

...multi-color imaging experiments. Interested in epitope tags? Others may be interested in attaching epitope tags... fluorescent proteins (FPs) to a yeast gene of interest. These fluorescently tagged genes are expressed...tags to their genes of interest, allowing for easy capture and detection of proteins and complexes, without...

...the first gRNA using restriction enzyme BbsI and the second gRNA using restriction enzyme BsaI. If you’... of gRNAs Golden Gate Assembly uses Type IIS restriction enzymes, which cleave outside of their recognition...These expression vectors each contain Type IIS restriction sites flanking the promoter-gRNA construct, but...oligonucleotides. These plasmids contain Type IIS restriction sites that flank the promoter-gRNA constructs...

...heavy restrictions on what I could and couldn’t do as an intern. I expected that these restrictions would...choose what gene to focus on. The gene choice had restrictions. It couldn’t be a Covid gene, a toxin, or an...running on a time crunch and knew that if we were to restart the procedure we would not have enough time. During...

... students to network, find groups with common interests, and settle into university life. The fairs are...1996, Cambridge Biosoc’s goal has been to promote interest in the biological sciences amongst the student...meetups. The society hopes to help young scientists interested in this field excel and develop their career ...from, and mentor while pursuing your research interests. Scientific societies like those mentioned above...

...well-equipped. Many institutions are sincerely interested in connecting with the public in a meaningful...scientific background and bring with them established interests and areas of expertise. Perhaps the strongest ... disparate spheres of research. In my case, an interest in neuroscience was carried over and allowed to... new intellectual environment. Over time, this interest came to incorporate other facets which occupied...

...plasmid with your gene of interest, you would replace mCherry with your gene of interest. pBLADEONLY: This plasmid...a pBAD33-based plasmid expressing your gene of interest, all you have to do is co-transform pBLADEONLY...pBLADEONLY. That way, a previously cloned gene of interest, under the pBAD promoter, can be controlled with light...

...available. Aguilar is a basic cell biologist. He got interested in Lowe Syndrome initially because of studies...disease associated with the gene, it piqued their interest. They obtained cells from patients with the condition...of the Wistar Institute, says that the lab got interested in XP out of a desire to understand the early...diseases at the same time. The limitation of budget restrained us from costly functional analyses.  “We believe...

...recapitulate the natural biology of a cell type of interest better than immortalized lines derived from the...it’s much more difficult to introduce a gene of interest into primary cells, so most primary cell lines...cell penetrating peptide (CPP) to a protein of interest thereby giving it the capacity to enter cells....29(1):33-41. PubMed. Protein transduction: unrestricted delivery into all cells? Schwarze SR, Hruska...

...standardized DNA constructs, but its use of Type IIS restriction enzymes results in scarless assembly. Finally...allowing fragments to be taken directly from restriction enzyme digests to DNA assembly. What makes it...containing regions of homology) cut from plasmids via restriction enzyme digestion, and used directly in DNA assembly...as easy as obtaining a plasmid with the gene of interest (search Addgene plasmids by gene here), designing...

...community by funding their travel to a conference of interest. In this post, we’re proud to announce our first...components - an enzyme that breaks down the HMO of interest into its component simple sugars (including lactose...controlled GFP. When the enzyme breaks down the HMO of interest, the lactose released activates expression of ...His-tagged ABC transporter known to bind to the HMO of interest. Fatima specifically proposes using this sensor...

...protein (BFP, GFP, or RFP) to the genomic loci of interest. Pairs of identical hairpins within one gRNA will...HaloTag with red fluorescent ligands to the loci of interest. Dual color detection In order to label two loci...sgRNA-Sirius-8XPP7) to recruit GFP to the loci of interest. To determine how the sgRNA-Sirius-8XPP7 and sgRNA-Sirius...that result in yellow fluorescence at the loci of interest, the team could visualize three loci and measure...

...Relationships need nurturing. Perhaps you’ve met someone interesting at an event. Or, perhaps you have a colleague...or other information that may be helpful or of interest to them. Don’t send a long email. If you have ...with people by discussing non-work topics and interests. Offer help when you can, instead of just asking... with people you don’t like or who do not show interest in you. Network with the people that you want ...

...site of interest. A gRNA is used to direct the PolI3M-nCas9 complex to a DNA site of interest, which nCas9...introduce diversification by targeting a site of interest with dCas9 fused to the Activation Induced Cytidine...nicking Cas9. A guide directs Cas9 to a site of interest, where it nicks its target DNA (left). Cas9 dissociates...

...with antibodies specific to your protein(s) of interest. Once the complex is captured, it is treated so...digestion can be appropriate when your protein of interest is a low abundance transcription factor or cofactor...point you have (1) crosslinked your proteins of interest to any DNA they were bound to (2) fragmented the...antibodies to specifically select for proteins of interest (4) isolated your antibody:protein:DNA complexes...

...your cloning strategy: Restriction enzyme digestion Be mindful of the restriction enzymes you chose. Make...which technique you are going to adopt (i.e. restriction cloning, Gibson Assembly, Golden Gate etc.), ... fragments of the size you expect, check the restriction pattern of the enzymes you chose, and verify ...

...plasmid, excluding any restriction enzymes or vector purchases (Table 1). Restriction enzymes and vector ...also a great way to get to know people doing interesting and relevant work in your field! However, there...work done previously, borrow small aliquots of restriction enzymes for one-off projects, or even save yourself...

...iterative one-pot reactions with straightforward restriction digestion and ligation steps to build barcoded...flexible and can accommodate any genetic elements of interest. It can thus be tailored to address the users’...effective gRNA sequences targeting your genes of interest. Thanks to the tremendous efforts made by various...the University of Hong Kong. He is particularly interested in developing and applying new technologies to...

...edition, we’re going to take a look at a very interesting tool that can be used for creating (excuse the...-lox” or “LSL” cassette) upstream of a gene of interest will prevent gene expression in the absence of...described above, a genetic modification can be restricted based on experimental constraints. This has been...used to successfully introduce Cre into cells of interest. What if I need two separate recombination events...

...transduced cells and loss of transgene expression (Interestingly, the strong immunogenicity of AdVs is what makes... in trans. The Gutless AdV have generated high interest for gene therapy due their increased cloning capacity...plasmids: a shuttle vector (in which the transgene of interest is cloned) and pAdEasy™ which contains the adenoviral...plasmid stock should be analyzed by endonuclease restriction digest or fully sequenced to confirm its integrity...

...inserting a targeting vector containing the gene of interest into a zygote, or into embryonic stem cells that...gene construct can be used to insert the gene of interest into a zygote or into embryonic stem cells. Non-homologous...genome using homology arms that flank the gene of interest. In either case, a positive selection marker such...the genome with similar homology to the site of interest or create mosaic animals where not all cells in...

...promoters could be an easier way to restrict expression of a gene interest in an animal model. A toolkit ...Applications for synthetic promoter AAVs Besides restricting AAV transgene expression to particular cell types...types, pairs of synthetic promoter can be used to restrict expression to a single cell type based on the ...

...vector file, and quickly clone their vector of interest. And it truly is plug-and-play. Because of its...recommend checking your cloning via a miniprep and restriction digest, and then plasmid sequencing if you can...Cloning specifics Fragmid uses Bbsl, a Type II restriction enzyme that cleaves outside of its recognition..., 93% had the correct banding pattern in the restriction enzyme digest. They sent 82 of the correctly ...

...since 2015 to conditionally degrade proteins of interest in cells (Zhang et al., 2015). Recently the Ward...degron (AID) sequence attached to your protein of interest. TIR1 interacts with endogenous Skp1 and Cul3 ...) tagged proteins. Degradation of your protein interest would also result in depletion of the FP, which...integrate the tagged AID* into genomic sites of interests via CRISPR techniques. This tool-kit includes ...

...finding journal articles related to your actual interests while also staying on top of the ever expanding...which is a nice way to see growth or decline of interest in a field over time. Creating an alert for a ...places to look for more articles on your topic of interest.  Best uses Accessing the vast body of biomedical...keywords, concepts, journals, and/or papers of interest. You can also just follow feeds curated by Meta...

... based off of p24 quantification tend to be overestimates since they can include free p24 and defective...assay, titering via measurement of viral RNA can overestimate the amount of infectious virus in a prep as ...physical titer, there is a risk that they may underestimate viral titer as these assays cannot distinguish...integration events, reducing the possibility of underestimating the viral titer. The main drawback of this ...

...transcribe, translate and modify your protein of interest. Modified proteins can then be extracted from ...is quite simple; DNA coding for your protein of interest is inserted into a plasmid expression vector that...propagate, are induced to produce your protein of interest, and then lysed. Protein can then be purified ...from Northeastern University and is particularly interested in cell signaling and communication. She loves...

...adjacent to the targeted sequence at the locus of interest. SpCas9’s 3’ NGG PAM occurs frequently in GC-rich...progress we’ve made, there is still a need for and interest in enzymes with shorter PAM sequences and/or broader...permits quick, large-scale mutation of the gene of interest. They created a selection phage (SP) to carry ...-3.13. To test xCas9 3.0-3.13, Hu et al. first restored catalytic residues D10 and H840. To ensure that...

...is to replace the genomic copy of your gene of interest with a fluorescent protein fusion. CRISPR genome...scientists to tag and light up endogenous genes of interest to better understand normal protein function. ...vectors:  1) a gRNA vector that targets the gene of interest, 2) a plasmid to specify the reading frame of ...flanked by long regions of homology to the gene of interest to promote homology directed repair after a CRISPR...

...that directs them to the promoter of a gene of interest, they have been shown to dramatically alter levels... transcriptional start site (TSS) of a gene of interest, it induces potent gene expression with no genetic...of the transcriptional start site of a gene of interest, Cas9 forms a steric block that prevents RNA polymerase...the length of gRNAs targeting different sites of interest thus represents a straightforward means of exerting...

...select for sybodies targeting a membrane protein of interest using the Sybody Generation Toolbox and commercially... (for purification purposes) protein target of interest, the protocol goes through 3 major steps to select...exchange (FX) cloning technique which uses Type IIS restriction sites (Geertsma, 2013). Once the phagemids are...bind strongly and specifically to the protein of interest. The Seeger lab used the ELISA technique described...

...allowing the reference sequence and the gene of interest to be amplified in the same reaction. Taqman’s...contains a primer and probe set for both the gene of interest and the reference sequence. At the end of the ...the number of positive droplets for the gene of interest and reference gene, copy number can be determined... your experiment?  Complexity of the gene of interest Some gene sequences are harder to amplify and...

... (5’) and downstream (3’) of the DNA region of interest. Primers are crucial for this process as DNA polymerases...times to exponentially replicate the target DNA of interest. This entire reaction used to be done manually...used in molecular cloning to create a plasmid of interest, for detecting the presence of an insert or piece...End point PCR or just PCR Amplify a region of interest  Molecular Cloning Site-directed mutagenesis...

...You're OK. However, if you fuse your protein of interest (POI) to GFP to study the protein's behavior in...reticulum (OSER) whorls. 2)A dimerizing protein of interest fused to a dimerizing FP has potential to form...protein concentrations. An imperfect solution is to restrict imaging analysis to cells expressing the lowest...Department of Anatomy and Structural Biology. His interests include the quality control of secretory proteins...

...initial reversal drive cannot restore the exact original sequence, it can restore the original protein-coding...using a recoding strategy; a subsequent drive can restore the wild-type sequence (save for the residual sgRNAs...immunizes it against the first drive. Laboratories interested in building candidate gene drives intended for...

...guide RNA (gRNA) targeting your genomic site of interest. This is commonly performed by transfecting cells...complex with a targeting gRNA, to your cells of interest. Cas9 RNPs are capable of cleaving genomic targets...CRISPR components to their experimental systems of interest. Using this method can alleviate many issues that...Hempstead is a Senior Scientist at Addgene.  Andrew's interests include genome engineering and microbiology.   ...

...fifth repeat Nobel Prize winner in history. Interestingly, he also adds to the disproportionately large...allowing every student to work on any topic of interest. The result was a positive experience for the ...generate new knowledge that will be of genuine interest to other researchers in their field. Indeed, sharing...Duboue et al., 2022), as they increase student interest in scientific career paths while also broadening...

... interaction trap), genes encoding proteins of interest(POI’s) are fused to TAs and DBDs (see Figure 2a...those cells where binding between the proteins of interest has occurred. Although the original Y2H systems...way, proteins that interact with a molecule of interest (the ‘bait’) can be ‘fished out’ for further study... random genomic fragments from the organism of interest, or a defined cDNA library (e.g. the entire proteome...

...technical challenges depending on their protein of interest. After several decades of trying to address these... protein? It is not generally in a cell’s best interest to overexpress a protein. Energy and cellular ...many tags and fusion proteins to your protein of interest would eventually get ridiculous (you generally...Life Sciences. Addgene pages and blog posts of interest: Which Fluorescent Protein Should I Use? Plasmids...

...but there are few genetic tools currently for terrestrial frogs. Cnidarian-Dinoflagellate symbionts (Virginia...food. They have chemical pathways generating interesting drug-like compounds especially cardenolides which...traits with good genetic mapping morphology and interesting behaviors. There is diversity in the social bonds...from other plants and are unusual enough to be interesting biologically. They have economic importance as...

... will your cells actually express your gene of interest? Many promoters are species-specific, so you’ll... to worry about the expression of your gene of interest in both systems you are shuttling between. For...probably don’t need it to express your protein of interest. However, you do still need a species-compatible...sometimes called transfer vectors. The gene of interest is cloned into a shuttle vector for transfection...

...If you’re interested in studying immunology or subpopulations of cells, you’ll soon find yourself encountering...markers to identify your specific cell population of interest. The markers you use, and the order you use them...like in Figure 3 above, until the population of interest has been successfully identified and isolated..../doi.org/10.5500/wjt.v8.i1.23 Jhunjhunwala, S., Aresta-Dasilva, S., Tang, K., Alvarez, D., Webber, M.,...

...inactivated native restriction/methylation system. This means the strain can neither restrict nor methylate...expression of T7 RNAP from the genome; does not restrict methylated DNA fhuA2 lacZ::T7 gene1 [lon] ompT...RNAP is then available to transcribe the gene of interest from a T7 promoter on the plasmid. Many commercial...