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...transferred from B. subtilis to the bacterium of interest following IPTG induction of conjugation genes ... transferred to a recipient bacteria strain of interest by tri-parental mating. Donor bacteria contain...ready to move Mobile-CRISPRi into your bacteria of interest? Find Mobile-CRISPRi plasmids at Addgene! References...

...with your transfer plasmid, carrying your gene of interest, and the two helper plasmids, commonly called ...Addgene AAV Titration by qPCR Protocol. If you’re interested in the ddPCR titering process, which is what ...non-genome-containing (empty) capsids, thus leading to an overestimate of transducible units in the virus preparation...

...tools allowed us to detect and capture proteins of interest to better study them in vivo and in vitro, especially... was to recombinantly append their proteins of interest with unrelated peptides that already had high ...only be added to the C-terminus of proteins of interest and proteins tagged with it needed to be chemically...

...pathway that are of interest to their laboratory. A single mouse click on a gene of interest will direct the...Michael G. Lemieux is a Scientist at Addgene with interests in education and scientific writing.     References...

...target near the last coding exon of your gene of interest. For ideal use, this gRNA should be cloned into...endogenous promoter and the 3’ UTR of the locus of interest, since the GFP-Puro will be inserted just upstream...nucleases and their applications. It will be interesting to see what new editing possibilities MMEJ can...

...have children but are interested in testing your SciComm skills or are interested in a career in education...Too complicated and I would quickly lose their interest. I won’t claim that I blew their minds, but I ...

...outcomes on viability if cells are arrested too long. Prolonged cell cycle arrest can lead to apoptosis in as... you don’t accidentally change your protein of interest.   Fig. 2: Stop re-cutting with PAM and...

... and efficient tools to introduce your gene of interest into cells. Unlike gamma-retroviruses that can...vector, a transfer vector including the gene of interest, and the plasmid coding for an envelope. Three...LTR promoter to drive expression of the gene of interest. Consequently, the levels of protein expression...

... this mutation by an adenine base editor would restore antibiotic resistance. To find such a protein, ...editing causes the creation or destruction of any restriction enzyme (RE) recognition site, RE analysis of ...University, University Park, USA. Dr. Molla is interested in precise genome editing in rice and uses base...

...discover new ways to reach our colleagues and other interested audiences quickly and concisely – video might...share any details about their research itself. Pinterest is also a favorite for millions and their visual...decide if reading more about the research is of interest. In his research, Scott Spicer concludes that ...

...isn’t for everyone, particularly if you’re more interested in selling a product than communicating your ...Reddit has a large and active community of people interested in science. You can see some of the conversations...prepared! The mechanics of doing the AMA were interesting. I started watching the questions and preparing...

...probability. Continuous inversion of the DNA sequence of interest can lead to poor expression of genes within that...lines that express Cre only in the cell type of interest (Atasoy et al. 2008; Cardin et al. 2009). Ligand...plasmids and the low concentration of recombinase restricts their expression to individual cells. With this...

...DNA or RNA nucleotides containing a sequence of interest (red) is amplified using Recombinase Polymerase...in complex with crRNA targeting the sequence of interest and an inactivated fluorescent RNA reporter. If...specifically targeted RNAs. Finally (and perhaps most interesting), Cas13a’s propensity to cleave any RNAs after...

...statistical methods, omission of some data, etc.  The firestorm that followed implementation of the policy comprised...Lastly, data is our raw product. If we don’t take interest in how it is released, we will be in danger of... publishers and funders. Clearly, it is in our interest to ensure that our data are reproducible and useful...

...necessary for getting the plasmid express our gene of interest. This element is responsible for initiating the...role in determining where and when your gene of interest will be expressed.      The RNA polymerase...results or even kill your cells if your gene of interest is toxic. To combat this, scientists have created...

...together as an interest group (15) While marchers are a group with one unifying interest (the advancement...thought of as just another party. Additionally, interest groups are often pandered to when strong or overlooked...

...activation motif (ITAM). By fusing ITAM to an RTK of interest and co-expressing a cytosolic ZtSH2 reporter, ...that pYtags can be generalized to other RTKs of interest, they create sensors for FGFR1, EGFR and ErbB2...KRaION potassium sensors Whatever your analyte of interest, it’s important to choose a biosensor that has...

...that’s what keeps things interesting. And hopefully they’ll be just as interesting as the finished product...partially reopened. But there were still many restrictions in place, most importantly, masks.  Besides ...

...Google.slides or Powerpoint file right now to capture interesting key results, striking images, creative ideas,... write itself when the time comes. 5. Choose interesting content Talk about something you know a lot about...commercialized, just talking about the possibility is interesting. Check out Joanne's Reddit AMA 6. Practice, practice...

...multiple exons that, when deleted through NHEJ, will restore the dystrophin reading frame. gRNAs bind in intronic...this result, Nelson et al. find that dystrophin restoration is maintained for at least six months. Future..., Olson EN. Postnatal genome editing partially restores dystrophin expression in a mouse model of muscular...

..., please refer to the paper. Why might one be interested in state machine technology? Biological research...and/or sortable reporters in certain states of interest. For example, in Figure 2 below we present a two-input...researcher in the Lu Lab at MIT. He is broadly interested in biocomputing and biological circuits.     ...

...able to knock-in a 2xFLAG epitope into my gene of interest, I encountered low efficiency (0.13%) and laborious.... Figure 1: In the background are 504 restriction digests of PCR products used to identify a single...CRISPR plasmids page. Future perspectives It is interesting to note that recipients for the 2015 Breakthrough...

...million results (Here’s the top one, in case you’re interested). “But I’m an introvert!” is not an excuse. Every...someone else on that team is putting their own self-interest ahead of the team’s goals, then that team is destined...other person really is acting against the team’s interests or not; once that seed of distrust is planted,...

...chemogenetic/optogenetic constructs into our cells of interest. Although they have a small packaging capacity...chemogenetics vs. optogenetics) I’m particularly interested in using chemogenetic and optogenetic AAV-based...2001): 1248. PubMed PMID: 11509958. 8. Ferrari, Forrest K., et al. "Second-strand synthesis is a rate-limiting...

...focus fluorescence from reaching a detector or restricting fluorescence to specific excitation volumes to...reduction in signal intensity. 2-Photon microscopes restrict excitation (and bleaching) to a single focal point... a large axial range. 2-photon imaging, which restricts excitation and photobleaching to the focal plane...

...transposase. The donor plasmid contains the gene(s) of interest flanked by terminal repeat sequences, which the...plasmids that express the reprogramming factors of interest. The advantages of this method are that it’s relatively...KLF4 LIN28 Check out these plasmids if you’re interested in Tet-inducible lentiviral expression of OSKM...

...2 - Getting high school students and teachers interested in JEI (the importance of word of mouth and personal...required for cytokinesis in JEI. Developing an interested in biology, working in a lab for the first time...

...Michigan State University with the backing of a prestigious Fulbright Fellowship. Dr. Kwapata now works as...genetics and biotechnology for Horticulture and Forest Crops at the Lilongwe University of Agriculture...

...move from microbiology to law? How did you get interested in pursuing law? BN: Life science was always ... general counsel at a small start-up is very interesting. It’s very varied. There’s IP work to do, general...

...and country. This information can be of personal interest to the depositor, but it can also be valuable ...annual reports. While the raw data is, of course, interesting to parse through, aggregation of this information...

...specificity allows scientists to target proteins of interest for labeling, quantification, purification, chromatin...the spleen of mice injected with the antigen of interest. The immortalized cells, called hybridomas, are...

...decision. In the open-access Data Hub, researchers interested in the Addgene antibody collection can look at... selecting an antibody! We encourage everyone interested in the Addgene antibody collection to use and...

...Even that grim estimate might prove to be an underestimate. Given all the excitement and speculation, we...ongoing outbreak from the CDC and WHO. - For anyone interested in EBOV’s past, present, and future, don’t miss...

...commercial kits you will adopt for the extraction. Restriction enzyme based approaches can require several hundreds...cloning grade DNA for direct cloning of your gene of interest into a plasmid backbone. Transfection grade...

...Citing Article" - such as cloning their gene of interest into a previously deposited vector - then you'...locate other papers that have used your plasmid of interest, which may help you identify relevant protocols...

...trainees at all. For grad students and postdocs interested in taking matters into their own hands, there...I am happy to work with any group of trainees interested in designing career exploration programs or peer...

... from your sequencing runs.   When you run a restriction digest on a gel you always include proper controls...primer at least 50bp upstream of the sequence of interest. 5. Use a silica spin column for purification ...

...to be used as research tools is particularly interesting. Although humans have known about bioluminescent...deciding which one is right for you, so if you’re interested in learning more about fluorescent proteins, ...

...this plasmid has a U6 promoter followed by BbsI restriction sites into which you can clone your 20 nucleotide...can use to express both SpCas9 and your gRNA of interest in a variety of mammalian cell lines. Alternative...

...Ubiquitin - One particular gene we saw increased interest in from 2008 to 2009 was ubiquitin. The most requested...next Top 10 list. 10 years of plasmid sharing  Interested in more facts about Addgene's 10 years of plasmid...

...can be applied non-invasively, its use is not restricted to cultured cells but extends to freely moving...plasmids have clear application to anyone with an interest in neuroscience, their use is by no means limited...

...discussed further in this informative post. Interestingly, this technology may make some uses of Site ... Software Engineer Kris Shamloo wrote a very interesting post about how we develop our in house lab management...

...screen for pathways that confer any phenotype of interest, not just drug resistance,” he said. “It could...contacted by investigators all over the world who are interested to use our libraries to do things - sometimes...

...characteristics and ectopic reporter assays. We have been interested in exploring the function of non-coding regulatory...frameshift mutations. However these may require restriction fragment length polymorphism analysis, allele-specific...

...double-stranded DNA break at the genomic loci of interest and on either side of the cargo (ex: fluorescent...the double-stranded break in the genomic loci of interest. As the universal sgRNA is not predicted to target...

...of AAV plasmids will have technical or legal restrictions that make them ineligible for use in this service...consideration will have a banner on their material page. Interested scientists can expand the banner to select their...

...Of course, if you’re interested in using IBEX, you’re probably also very interested in having high-quality...

...activated in response to a ligand binding a GPCR of interest. How does TRUPATH work? The TRUPATH collection...transfected with plasmids expressing the GPCR of interest, a Gα-Rluc8 plasmid, and the appropriate Gγ-GFP2...

...between recurring meetings, you should share interesting results or ask questions to your collaborators...microscopes and allowed me to investigate my protein of interest in a French clinical cohort, neither of which ...

...specifically isolate DNA that is bound by a protein of interest. First, nuclei from tissue or cell culture are...with a primary antibody against the protein of interest. The Protein A fused MNase is then added and Protein...