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We narrowed to 754 results for: REV

Showing: 121 - 160 of 754 results
  1. Antibiotic Resistance: An Old Solution but a New Problem

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    Blog Post
    ... any preventive measure designed to help keep you from getting sick can be helpful in preventing the spread... on preventing the spread of antibiotic resistant bacteria, check out this resource on preventing antibiotic...because ‘antibiotic’ merely means that the drug prevents the growth of a living organism. As for multi-...standpoint, we can all work together to try and prevent outbreaks of drug resistant bacteria within communities...standards. Handwashing is an essential component of preventing infection, as is cleaning and disinfecting frequently...
  2. Rett Syndrome: A History of Research and Therapeutic Outlooks

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    Blog Post
    ...individuals with Rett syndrome experience regression of previously acquired skills such as speech, coordination,...landmark paper in Science showed that dramatic reversal of disease symptoms in mice is possible (Guy et... halted the neurological damage but completely reversed it, regardless of age or symptom severity. This... exciting finding injected hope that a similar reversal may be possible for children and adults with Rett...RNA trans-splicing, and protein replacement to reverse the impacts of this disease.  As a non-degenerative...non-degenerative, monogenic disease that has shown reversibility of symptoms in animal models, Rett syndrome is ...Guy J, Gan J, Selfridge J, Cobb S, Bird A (2007) Reversal of Neurological Defects in a Mouse Model of Rett...
  3. 28 Hot Plasmid Technologies from 2015

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    Blog Post
    ... and the promoter to prevent leaky expression of this resistance. Unlike previous systems, pUCXKT does...can do so in a very specific, precise manner. Previously, scientists have been successful in regulating...enzymes (e.g. nucleases, recombinases) which irreversibly excise or invert a specific chunk of DNA. However...deposited a set of 11 phage integrases which act irreversibly on their cognate attB/attP sites with no cross-talk...subcellular localization in mammalian cell culture and reversible control of signalling pathways. These LIDs are... powerful tools which allow you to easily and reversibly switch off and on your favourite signalling pathways...the MCS has several options) and a pUCX-specific reverse primer containing the missing codons from the antibiotic...
  4. FLEx Technology and Optogenetics: Flipping the switch on gene expression with high spatial and temporal resolution

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    Blog Post
    ... have taken researchers to the threshold of a revolution in modern neuroscience, an era in which technologies...antisense orientation with respect to the promoter to prevent its expression, and is flanked (“floxed”) by two...electroencephalography evidences of arousal in mice previously subjected to steady-state general anesthesia ...behaviors. The high spatio-temporal precision and reversible modulation of FLEx vectors combined with the ...functions, and molecular mechanisms." Chemical reviews 114.1 (2013): 126-163. PubMed PMID: 24364740. PubMed...development and application of optogenetics." Annual review of neuroscience 34 (2011). PubMed PMID: 21692661...of cre-loxp site-specific recombination." Annual review of biophysics and biomolecular structure 30.1 (...
  5. Hot Plasmids - September 2019 - CRISPR, Auxin Inducible Degradation, MerMAID Channelrhodopsins, and Genome Looping

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    Blog Post
    ...plant hormones called auxins to conditionally and reversibly degrade proteins of interest fused to a degron...2009, Natsume et al., 2016).  The Kanemaki lab previously developed a CRISPR/Cas9 based method to tag endogenous... be used to engineer specific sites that were previously difficult. Find SpCas9 and Target-AID-NG plasmids...promoter and an enhancer. What makes LADL unique from previous iterations is the addition of CRY2, another Arabidoposis...hours of blue light induction (6-fold faster than previously developed systems). The LADL system promises ...it’s briefer time scale and its inducible and reversible nature. Find the LADL plasmids at Addgene Kim...
  6. Plasmids 101: The protein expression toolbox

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    Blog Post
    ...Turn it on? Turn it off? Turn it down, but not forever? You don’t need to resort to the force to control...biological tools are abundant! In this post, we will review tried-and-true protein expression control tools...the options together. Turn it off Turn it off forever If you want to turn off protein expression, you...for transient knockouts!  Turn it on Turn it on forever Permanently increasing or turning on expression...amounts of tetracycline. Tetracycline regulation is reversible (when the drug is removed expression goes back...doesn’t require this level of control, one of the previous options may be better suited for the job. Whether...
  7. A Look at Addgene's QC Process

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    Blog Post
    ... issue is reviewed yearly at a best practices meeting, where the QC team updates and reviews our sequencing...caught prior to distribution. Catching these errors prevents future experimental issues, saving time and money...encountered in plasmids deposited to Addgene. This review process also highlights other QC topics that may...team suspects an assembly issue, they manually revisit the raw NGS reads and reassemble using a program... issue, they will contact the depositing lab to review the discrepancies. Often, the identified issues...
  8. Design Tips for Prime Editing

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    Blog Post
    ...that relies on a fusion enzyme of Cas9 nickase + reverse transcriptase and a prime editing guide RNA (pegRNA...binding site (PBS) complementary to the genome and a reverse transcriptase (RT) template including the desired...Anzalone et al., 2019). Test different lengths of reverse transcriptase template, starting with about 10–...optimizing your pegRNA primer binding site (PBS) and reverse transcriptase template (RTT). Created with BioRender.com...Edit the PAM along with your intended edit. This prevents the Cas9 nickase from re-binding and nicking the... al., 2019). Figure 4: Editing the PAM prevents the prime editor from re-engaging with DNA it ...
  9. How to Deposit Your Plasmids with Addgene

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    Blog Post
    ...is published. Plasmids can be linked to a peer-reviewed manuscript OR a preprint. In the case of a preprint...information once the manuscript is published in a peer-reviewed journal. To start this process, simply enter your...Step 5: Verification On this page you will see a preview of the final plasmid page that will appear on Addgene...once your plasmids are available online. Please review all of the information on this page to make sure...easily update the information by navigating to the previous pages using the links at the top of the page. .... Figure 11: A completed plasmid page for review before submission. Once you are satisfied with the plasmid...backbone, you can copy some information from a previous plasmid in your deposit by selecting the plasmid...
  10. Transgenic Organisms, Cas9 Gene Drives, and Appropriate Safeguards

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    Blog Post
    ...one mistake can be enough. Reversal drives are designed to overwrite a previous gene drive and thereby undo...by the earlier intervention2. While an initial reversal drive cannot restore the exact original sequence... sgRNAs and possibly cas9 gene). An immunizing reversal drive is a variant that also spreads through the...should consider building an appropriate immunizing reversal drive at the same time to mitigate the potential...M. RNA-guided gene drives can efficiently and reversibly bias inheritance in wild yeast. bioRxiv 013896...
  11. The CRISPR Software Matchmaker: A New Tool for Choosing the Best CRISPR Software for Your Needs

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    Blog Post
    ...fill the prop with something of value. They are reviewers of content. On the other hand, the goals of the...defined by their trust in the piñata builders - the reviewers. After some physical exertion on the part of the... the value of each item changes from what the reviewers so painstakingly assessed to something a bit more...”, it is the disconnect arising between those reviewing the contents of the piñata and the audience blindly...software development has caught up with it. The peer review process has been challenged with many closely spaced...variations on a theme without many resources to review them. Collectively, the pool of CRISPR software...view and get a sense of which sgRNA designs have previously worked and under what conditions. Such resources...
  12. A Career in Grants: How To Become a Grant Professional

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    Blog Post
    ... to review grant proposals from others. Dr. Cook was asked to serve on review panels and to review education...profession? I asked Deborah Cook, Ph.D., GPC, previously a college biology professor, how she became a...development? Volunteer as a grant writer and/or grant reviewer with a local nonprofit that aligns with your passion...Profession. Whether you’re transitioning from a previous career or you’re starting your first career, a...
  13. CRISPR Activation: A Practical Guide

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    Blog Post
    ...activating genes that cause differentiation and to reverse HIV viral latency (Bialek et al., 2016). Additionally...2015). These types of CRISPR activation are also reversible: once the environmental trigger leaves, gene ...activated. However, there are several factors that may prevent your gene from being up-regulated. For instance...This is believed to be true given the results of previous RNA-seq experiments (Konermann et al., 2014, Chavez...Hauber J, Lange UC (2016) Targeted HIV-1 Latency Reversal Using CRISPR/Cas9-Derived Transcriptional Activator...10.1016/j.cell.2014.09.029 Konermann S, Brigham MD, Trevino AE, Joung J, Abudayyeh OO, Barcena C, Hsu PD, Habib...
  14. Advanced Uses of Cre-lox and Flp-FRT - A Neuroscientist’s View

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    Blog Post
    ...invert or ‘flip DNA’, is derived from the yeast S. cerevisiae (Gronostajski and Sadowski 1985). The FLP system...recombination events mediated by FLP or Cre are reversible. Whereas the excision of a piece of DNA flanked... within that sequence and should therefore be prevented. The loxP and FRT target sites have been engineered...sites (Schnütgen et al. 2003). Flex vectors revolutionize neuroscience Manipulation of specific cell types...channels (plasmid # 20298 for example) have revolutionized Neuroscience. They are the only means to target...1985. “The FLP Recombinase of the Saccharomyces Cerevisiae 2 Micron Plasmid Attaches Covalently to DNA via...
  15. Using Phosphoserine to Study Protein Phosphorylation

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    Blog Post
    ...suggest that phosphorylation is far more prevalent than previously appreciated. PhosphoSitePlus, one of the... group. Additionally, these mutations are not reversible, and thus cannot reflect the true state of protein...Extensive genomic sequencing of prokaryotic lineages revealed a curious anomaly amongst a subset of methanogenic...mutation of these residues. Due to the limitations previously mentioned, phosphomimetic mutations on the activation...affects protein function, researchers should try to reverse the effects of pSer-incorporation on enzyme function...
  16. Hot Biosensors 2022: Year-End Roundup

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    Blog Post
    ...photostability under two-photon illumination than previous GEVIs. Some constructs are also available as viral...kinetics and photochemical characteristics than previous NIR calcium reporters iGECI and NIR-GECO2G, and...reduced sensitivity to pH and calcium compared to previous lactate biosensors, and its affinity is better-matched...temperature sensor that significantly outperforms previous designs. B-gTEMP is excitable under blue light...B-gTEMP itself is a significant improvement over previous temperature indicators, it also demonstrates that...Intracellular Heat Transfer and Thermal Property Revealed by Kilohertz Temperature Imaging with a Genetically...
  17. Evolution of Brainbow: Using Cre-lox for Multicolor Labeling of Neurons

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    Blog Post
    ...the continued utility of Cre-lox. Check out our previous blog post, Plasmids 101: Cre-lox, if you need ... removes one of the other loxP site variants, preventing further recombination. Brainbow-1.1 scales up...that transient Cre recombination is required to prevent constant inversion of the construct. Sanes and...their protein sequences are highly homologous, preventing design of antibodies specific to each fluorophore...of delicate axonal and dendritic processes not previously visible with Brainbow-1 and -2. The general structure...connectome. Lichtman JW, Livet J, Sanes JR. Nature Reviews Neuroscience. 2008 Jun;9(6):417-22. doi: 10.1038...
  18. Addgene's a Nonprofit? Nonprofit Awareness Day 2017

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    Blog Post
    ...as a nonprofit under the United States Internal Revenue Code 501(c)(3). That means we were formed to benefit... are able to fulfill our mission by generating revenue through distribution of research materials. Our..." is a bit misleading. Nonprofits can generate revenue (as Addgene does through materials distribution... nonprofits should have some level of positive revenue to build reserves so the company is sustainable...The difference is what nonprofits do with their revenue. A nonprofit organization cannot distribute profits...
  19. How Dry Ice Affects Viral Vector Sample pH & How to Avoid It

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    Blog Post
    ... news is that this effect is reversible and the pH shift can be prevented. Keep this information in mind...if you’re receiving samples on dry ice - it may prevent you from seeing some unexpected results. Order... 3). However, 30 seconds was not sufficient to prevent any pH change, as we still observed a slight change... allowed the CO2 to leave the tubes, and would prevent the pH change upon thaw (Fig.4). To test this, ...
  20. Management for Scientists: Delegating is Key

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    Blog Post
    ...entirely too much work for one person, it would take forever, and the end product would likely be poor quality... delegate the giving of feedback or performance review. You can’t delegate tasks assigned to you and you...outcomes, timelines and deadlines. Agree on a way to review project progress. I recommend putting a regular...approachable mistakes will be made that could have been prevented.  Make sure everyone understands that no question... Aren't You Delegating" in the Harvard Business Review. Another excellent blog on delegation tips and ...
  21. CRISPR 101: Cytosine Transversion Editors

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    Blog Post
    ...your vocabulary! A brief review of base editing tools First, let’s briefly review how CRISPR-derived base...The first base editors revolutionized CRISPR gene editing. Cytosine base editors (CBEs) and adenine base...the pyrimidine bases (C and T). (Check out our previous post on these editors for more details on how ...also called apurinic or apyrimidinic sites and abbreviated as AP sites. To explore this further, the team...bound, keeping the two DNA strands separate and preventing repair based on the complementary strand. This...
  22. Hot Plasmids December 2018 - C. elegans RNAi, TurboID, Detecting Viral Outbreaks, and Protein Binding Specificity Tools

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    Blog Post
    ...sequencing, revealing that they bind to a much larger subset of protein motifs than previously thought. ...T7 terminators adjacent to the T7 promoters to prevent the incorporation of non-specific nucleotides into...proteomics in model organisms in which it was previously not an option, including Drosophila and C. elegans...Pubmed PMID: 30370338.  SH3 domain collection reveals novel peptide ligand motifs Article contributed...
  23. Choosing the B(right)est Fluorescent Protein: Aggregation Tendency

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    Blog Post
    ...Advanced microscopy, University of Amsterdam. The previous two posts in this series described a practical...between the FPs interfere with native interactions, preventing proper localization in these assays. However,...tendency in living cells. Strikingly, the OSER assay reveals that tagRFP does not act as a true monomer in cells... cells (Constantini, 2012), whereas it was previously concluded to be monomeric based on gel filtration... that are relevant for your research. This will reveal the performance of fluorescent proteins in your...
  24. Writing Scientific Manuscripts: Literature Searching, Reading, & Organizing

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    Blog Post
    ... titles, authors, year, narrow by article type (review, patent, clinical study, research article etc)....on a new literature search, try to find a recent review article to give you an overview of the topic and...information dense and since it continually builds on previous studies, the reader always enters in the middle...introductions should be fairly focused. Consult a review article for a broader scope of what’s going on ...performed that could influence the results? The peer-review process is not perfect, so even though it is science...
  25. Deaminet 2026: Breakthroughs in Base Editing, Deaminase Biology, and Therapeutic Translation

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    Blog Post
    ...comes with a trade-off: overextension past the reverse transcriptase (RT) template, resulting in unintended...Structural data implicate specific residues in the reverse transcriptase polymerase domain as likely determinants...Riverside) presented molecular simulation studies revealing that TadA8e dimerization, together with a unique...off target sites than edit-only approaches and revealed distinct noncoding off-target landscapes for ABE8e...In clinically relevant HSPC models, Inrich-seq revealed numerous high-frequency off-target sites in the...AAV-delivered adaptamer-controlled FGF21 expression reversed obesity and metabolic comorbidities, demonstrating...
  26. Rabies and Neuronal Tracing

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    Blog Post
    ...inputs. This study revealed that these neurons received more inputs than was previously thought. The differences...subsequently higher and higher order neurons doesn’t reveal network connections in a useful way, but rather... tracing is a powerful tool that can be used to reveal brain circuitry, and continue to contribute answers..., Berninger B. Retrograde monosynaptic tracing reveals the temporal evolution of inputs onto new neurons...
  27. CRISPR 101: Anti-CRISPR Proteins Switch Off CRISPR-Cas Systems

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    Blog Post
    ...In this study, the labs found that anti-CRISPR prevents Cas9 activity in HEK293 by reducing editing from... Acr samples. Acr proteins can also be used to prevent dCas9 tethering to DNA. Since these early anti-... AcrIIC2 interactions with Cas9 interferes and prevents the correct assembly of the crRNA-Cas complex ...binds to the HNH endonuclease domain of Cas9 and prevents target DNA cleavage (Harrington et al., 2017)....vectors. Having a way to shut off CRISPR-Cas systems prevents the off-target effects and cytotoxicity that result...
  28. CrispyCrunch: High-throughput Design and Analysis of CRISPR+HDR Experiments

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    Blog Post
    ...explosion of software tools accompanying the CRISPR revolution. (See omictools for even more.) However, none...step of mutating guide sequences in a way that prevents re-cutting but does not disrupt gene expression...CrispyCrunch is how it mutates guide sequences to prevent re-cutting of the sequence inserted by HDR. Although...mutating a single base pair may not be enough to prevent re-binding. Lastly, the sequence inserted by HDR... target regions Design donor RNA Design primers Review and order reagents Perform your wet lab experiment...
  29. Deep Dive: qPCR

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    ...and I recommend Bustin, 2017 for a more in-depth review.)  Your amplicon should be between 70-200 bp long...ideally located close to either the forward or the reverse primer.  There is a list of useful primer design...efficacy with a standard curve. If relying on previously-generated validation data, be aware that different...transcripts. However, adding too many reagents can prevent your reaction from reaching the plateau stage after...sample compared to your control. But as we said previously, just because it’s used often doesn’t mean it...York 2014Bustin S; Huggett J. qPCR primer design revisited. Biomolecular Detection and Quantification. Volume...
  30. Deep Dive: Fixing and Permeabilizing for Immunofluorescence

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    ...thus structural epitopes, but can reveal linear epitopes previously hidden within the protein. Created...cross-linking can block the epitope of some targets, preventing your primary antibody from binding (Fig.1). The...cleaner results by not permeabilizing as you’ll prevent binding to any target protein that is still on ...preserve sample morphology, while organic solvents can reveal otherwise hidden epitopes. However, keep in mind...
  31. Plasmids 101: Stringent Regulation of Replication

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    Blog Post
    ...always to the same extent. As described in our previous Origin of Replication post, DNA replication is...bacteria Before looking at an example plasmid, let's review how E. coli normally replicate their chromosomal...sequence forming this pseudoknot, so its binding prevents the mRNA from folding and blocks RepZ translation...high levels of the lncRNA bind to RepZ mRNA and prevent this pseudoknot from folding, blocking replication...bacterial plasmids. Microbiology and Molecular Biology Reviews: MMBR, 62(2), 434–464. https://doi.org/10.1128/...
  32. Deciphering the Mysteries of Behavior with Viral Vectors

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    ... unraveling the mysteries of behavior. We’ve previously discussed some of the general considerations ....  What’s involved in locomotor sensitization? Previous research has shown that regions such as the nucleus...transmission leads to locomotion sensitization. (for review see van Huijstee & Mansvelder  2014). Scientists...the locomotion of the animal. This confirms the previously suggested inhibitory mechanism of locomotor sensitization... times are ahead for behavioral neuroscience to reveal insights into the complex relationship between ...
  33. Advancing Biology with Zebrafish: Genetic Tools for Developmental Studies and More

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    ...experimental treatment can be monitored. This can reveal previously unknown functions for genes, and lays the ...of the zebrafish genome was published in 2013, revealing that about 70% of human genes have a counterpart... ways. Zebrafish mutants In the 1990’s, two revolutionary large-scale random mutagenesis screens produced...unlocked novel research into genetic compensation, a previously unknown mechanism of gene regulation (El-Brolosy...
  34. Data Freedom: The Expansion of Data Sharing in Research Publications

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    ...publications. The policy was not an overnight revelation, rather it was the result of consultations between... rationale includes increasing the ability of reviewers (and, once published, anyone) to detect possible...apply different or novel tools and assumptions to reveal findings that the originator was oblivious to, ...called Scientific Data, which aims to act as a peer-reviewed repository for “scientifically valuable” datasets...or table, clicking on the published chart could reveal the underlying data and allow other analyses “on...
  35. Transferable Skills Guide: Time Management

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    ...s post and give it a final read Browse list of previous depositors from Boston University Contact Dr. ...that post from Eric before doing anything else. Preventing procrastination Even if you’ve done all the prioritizing...planning in the world, procrastination can easily prevent you from making progress on a task. For instance...to go sit in the tissue culture hood may have prevented me from splitting cells until late in the day ...podcasts. JUST GET STARTED - One Harvard Business Review article on procrastination recommends testing out...
  36. CRISPR 101: Homology Directed Repair

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    ...Saccharomyces cerevisiae. Paques F and Haber JE. Microbiology and Molecular Biology Reviews (1999). PubMed...repair and genome engineering CRISPR-Cas9 has revolutionized the genome engineering world and made targeted... disrupting one or both of these sequences. To prevent re-cutting and the resulting mutations after an...transcribes RNA encoding your repair template, then uses reverse transcriptase to generate the complementary ssDNA...
  37. SARS-CoV-2/COVID-19 Detection Methods Based on CRISPR/Cas

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    ...gene are amplified. RPA reactions also contain reverse transcriptase to produce cDNA. Isothermal amplification...are designed, each binding to one strand of the reverse-transcribed target sequence that are present close...with 9V battery Approach :This preprint uses a previously developed method to detect SARS-CoV-2 in the ...samples. First, RNA is extracted and subject to reverse transcription. This is then amplified by PCR using...detect fluorescence upon cleavage. Highlights: Reverse-transcribed RNA samples are amplified by PCR instead...
  38. Thoughts on the Future of Research 2015 Boston Symposium

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    ...lack of complete information does not, however, prevent us from identifying problems with the structure...take on opportunities to do some blogging, write reviews, or write for a university publication, improving...was on diversity in academic research. As during previous panels, speakers Moon Duchin, Rafael Luna, Joan... need some way of capturing, quantifying, and preventing this type of failure. Final thoughts The FOR ...Additional Resources The Postdoctoral Experience Revisited. PubMed PMID: 25590106. Paula Stephan’s Book -...
  39. Easi-CRISPR: Generating Knock-In and Conditional Mouse Models

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    ...efficiency of 30-60%. This is much more favorable than previous methods yielding 1-10% knock-in. Read on to learn...transcription, and use the IVT product as a template for reverse transcription to create your ssDNA. Note that the...conditional deletion of exon 6 of Syt1, mimicking a previously made mouse model. To avoid disrupting splice ...separates the gRNA target sequences from their PAMs, preventing re-cutting after editing. How to use Addgene ...Additional Resources on the Addgene Blog Read Our Previous Post on CRISPR Mouse Models Learn about Homology...
  40. Antibodies 101: Antibody Engineering and Directed Evolution

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    ...applications. Chemical Reviews, 121(20), 12384–12444. https://doi.org/10.1021/acs.chemrev.1c00260 Wilson, D...quicker than others. We’ve covered some methods in previous blog posts, like developing recombinant antibodies...antibodies for autoimmunity and inflammation. Nature Reviews Immunology, 10(5), 301–316. https://doi.org/10.1038...for the directed evolution of proteins. Nature Reviews Genetics, 16(7), 379–394. https://doi.org/10.1038...
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