...is currently investigating the molecular biology of synaptic transmission and plasticity.     Javier García-Nafría...co-transformation with linearized vector backbone. DpnI digestion - 1 μl DpnI is added to the PCR mix to remove ... García-Nafría is an investigator scientist at the MRC Laboratory of Molecular Biology. He is interested...

...-wide screens to effectively and efficiently investigate gene function. CRISPR libraries can be used to...labs of David Root and John Doench at the Broad Institute have developed human genome-wide CRISPRko (knock...lowly expressed genes or genes that are more drastically affected by gene activation rather than depletion...resistance in A375 cells. The SAM system (CRISPR/Cas9 Synergistic Activation Mediator) uses an improved activator...

.... To kick off a knock-in experiment the first questions are where, which, and what. Identify where in ...like to introduce the edit. Finally, you must stipulate what sequence to introduce, which is done through...HDR edit, you can try one or more of the below suggestions. Disable the other repair mechanisms Mammalian...! Cas12a is a Cas family member which generates sticky ends upon cutting. Cas12a may promote HDR by generating...

...Janelia Research Campus, Howard Hughes Medical Institute. Chemistry is dead, long live chemistry! The discovery.... These impressive advances lead to an obvious question: In this age of GFP and its ilk, why should cell...Janelia Research Campus, Howard Hughes Medical Institute. He would rather be in the lab making new fluorophores...PubMed PMID: 26575238. 5. Crivat, Georgeta, and Justin W. Taraska. "Imaging proteins inside cells with...

...bit of stochastic magic and Poisson statistics, each sub-population has, on average, one distinct enhancer...intuition for how genome math helps us to locate interesting addresses in the genome. Let’s now examine how...

... vector (deposited by David Root of the Broad Institute) has consistently been one of the repository’s...have used the protocol for this plasmid, and if it still being used. After learning of it's continued popularity...is the creation of ZappyLabs. Currently in beta testing, the software aims to be a life science protocol...bring your phone in the lab, though, honestly, this still grosses me out. Maybe it’s just me – I didn’t get...

...methods measure different characteristics of the solution. These characteristics give us information on ... tries to get the other to guess a category by listing off things that fall into that category? Okay, ...do our best to send you a particular lot (if we still have it available). References 1. Fagone, Paolo...

...with a different editing window than existing CGBEs. This distinct approach means researchers can now select...the adenine transversion editor AYBEv3. This tool still retained considerable A → G editing activity at ...rates ranging from 4.7% to 30% at tested sites. Still, it opened a new avenue for base editor development...

...same blind spots, and there wouldn’t be anyone to question these blind spots. Blind spots often result in...collaborates with stakeholders to resolve the questions.  After the code is written, it is reviewed by...these libraries as ingredients to a recipe. It’s unrealistic to plant, grow, and harvest grains every time...news Resources on Addgene.org Check out our job postings Learn more about Addgene's mission ...

...cardio-respiratory dysfunction, distinctive hand movements, and show autistic-like behaviors.  The role of...animal models results in rapid symptom onset, suggesting that MECP2 is also essential after development...). Beam Therapeutics, co-founded by Harvard Investigators and Addgene depositors David Liu, PhD, Keith...

... delve deeper and look specifically at the characteristics and capabilities an ELN needs to facilitate...Goettingen, and several other large research institutions. The following graphic depicts the ways in which...banks and sequencing files is likely stored in institutional and lab servers specifically designed to hold...illustrated in this video, and to data stored on institutional and lab servers. Connectivity to other tools...

... PMC5726555. 4. Xie, Kabin, Bastian Minkenberg, and Yinong Yang. "Boosting CRISPR/Cas9 multiplex editing... a single base. If you heard about CRISPR, one question probably comes to mind: can CRISPR be applied ...Ag-biotechnologist in the National Rice Research Institute, Cuttack, Odisha, India. Currently Kutubuddin ...

...there are still many more features to consider when designing a full gene. A non-exhaustive list includes...be many possible ways to encode them and what constitutes the “appropriate” codon is not necessarily obvious...PubMed Central PMCID: PMC2970903. 6. Pechmann, Sebastian, and Judith Frydman. "Evolutionary conservation...

...Understanding the characteristics of natural plasmids and how they function in nature can inform us on...other hosts. For example, the pJD4 plasmid harbors distinct origins (ori1, ori2, ori3) and replication initiation...these plasmids are compatible with.  Table 1: Non-exhaustive list of common broad host range plasmids and ... P. C., & Wong, H. L. (2023). Development of constitutive and IPTG-inducible integron promoter-based expression...

...the Wyss Institute for Biologically Inspired Engineering. CRISPR/Cas9 is an enormously plastic tool and... University and former researcher at the Wyss Institute for Biologically Inspired Engineering.     Alejandro....2014.09.039 Wong N, Liu W, Wang X (2015) WU-CRISPR: characteristics of functional guide RNAs for the CRISPR/Cas9...

...interneurons across species and will undoubtedly constitute a valuable tool for the research community. You...CaMKII and RhoA during induction of structural plasticity in single dendritic spines. The set of CyRFP ...variants to develop five new base editors with distinct PAM sequences, expanding the number of available...xylose-inducible dCas9 paired with a single, unique, and constitutively expressed sgRNA targeting the 5’ end of each...to premature aging and cancer. The Denchi lab investigated an aspect of telomere length regulation called...

... It can be so overwhelming that you might just stick with the protocol your labmate handed you — after... contain any denaturing agents. However, you’ll still have to lyse your proteins. For this post, we are...stronger reducing agent than BME and has a less… distinctive smell. BME smells like rotting eggs but has a...However, these systems are expensive, generate plastic waste, and limit the amount of optimization that... may only have access to one or another. Most institutes have moved towards CCD cameras over the past ...

...membrane to enter cells. Depending on the research question and downstream application, one or several of ...are protected. Amine-binding viability dyes can still bind free amines on the cell surface, which yields...As a result, the largely negative live cells can still be easily gated on during the analysis. Conclusion...Harvard Medical School. His projects include investigating negative regulators of T cells and cytokine ...

...customer support, but there are eight other teams who still retain a customer support element amongst their ...manager to explicitly assign the responsibility in question to a specific person or team. What Makes You ... will not be great at resolving conflicts.  Institutional Knowledge: Of the tools I’m mentioning here,...have used your empathy, listening skills, and institutional knowledge to resolve conflicts. Conflict in ...

...cellular world. The technique allows you to ask questions like: “Where does my protein of interest live ...corresponding secondary antibodies conjugated to distinct fluorophores to visualize multiple targets at ...Even with all of the resources available, it may still be difficult to find an antibody in which you are...not only answers to your most pressing research questions, but also stunning images (am I the only one with...