We narrowed to 841 results for: STI
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TypeBlog PostPublishedOct. 23, 2018, 11:49 a.m....Restriction digest screening for ITR deletion is still necessary. To save time and money, I like to miniprep...Production protocol, I take a lazier approach for harvesting my AAV. I skip the PEG precipitation of the HEK293...PEG solution. PEG takes several hours on a heated stir plate to dissolve and requires monitoring so that...autoclaving, which also takes time. Rather than investing all of this effort in preparing reagents, I choose...
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Tips for Using BLAST to Verify Plasmids
TypeBlog PostPublishedMay 29, 2014, 1:29 p.m.... the “Align two or more sequences” option to drastically reduce waiting time for search results. Optimizing...with the "Align two or more sequences" option and pasting your expected protein sequence into the Subject...webservices/#pairwise_sequence_alignment_psa Happy BLASTing! Do you have any tips for using BLAST to confirm...plasmid sequencing results or comments on our suggestions? Share your thoughts here to help other labs ... -
CRISPR 101: RNA Editing with Cas13
TypeBlog PostPublishedJuly 31, 2020, 12:30 p.m....editor, the lab started with the Cas13 scaffold, testing a whopping 21 Cas13a orthologs, 15 Cas13b orthologs... the plasmids for the RESCUE RNA editors here! Testing and improving RNA editing from the Cas13b REPAIR...knockdown is more precise than shRNA knockdown, it still displays substantial off-target activity. By mutating...Addgene. East-Seletsky, Alexandra, et al. "Two distinct RNase activities of CRISPR-C2c2 enable guide-RNA... -
Antibodies 101: Producing Recombinant Antibodies
TypeBlog PostPublishedAug. 13, 2024, 1:15 p.m....2020). Harvest and purification At its core, harvesting recombinant antibodies consists of separating...Created with biorender.com Another method of harvesting uses magnetic beads. In this method, beads are... the efficacy of our antibodies through stress testing. An aliquot of our recombinant antibodies is stored...assays showed that the majority of antibodies are still equally effective compared to those aliquots frozen... -
CRISPR 101: Making a Knock-In Cell Line
TypeBlog PostPublishedDec. 22, 2022, 2:15 p.m..... 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... -
Introducing Addgene's New Search - Find Plasmids More Easily For Your Research!
TypeBlog PostPublishedDec. 3, 2019, 11:38 a.m....clicking the ‘Search’ button a new content-aware suggestion dropdown list will let you jump directly to the...types of searches including depositing labs and institutions. For example, search for “Melina Fan” and you... in Parkinson’s disease. Knowing the primary investigator where the plasmid was created can narrow your...plasmids specifically designed for this purpose: Still stumped? Ask an Addgenie The newly launched search... -
Simplify Cloning with in vivo Assembly
TypeBlog PostPublishedOct. 18, 2018, 12:37 p.m....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... -
New Optimized Genome-wide CRISPRko, CRISPRi, and CRISPRa Libraries
TypeBlog PostPublishedOct. 4, 2018, 12:44 p.m....-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... -
Better Dyeing Through Chemistry & Small Molecule Fluorophores
TypeBlog PostPublishedSept. 8, 2016, 2:30 p.m....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... -
CRISPR Between the Genes: How to Experiment with Enhancers and Epigenomics
TypeBlog PostPublishedAug. 9, 2016, 2:30 p.m....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... -
Sharing Your Lab Protocols: Using Apps to Save Time & Track Your Experiments
TypeBlog PostPublishedSept. 23, 2014, 3:05 p.m.... 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... -
15 Hot Plasmids from 2017
TypeBlog PostPublishedJan. 9, 2018, 3:02 p.m....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... -
Technical Design of a Western Blot
TypeBlog PostPublishedAug. 22, 2024, 1:15 p.m.... 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 ... -
Supporting Reproducibility with a Connected ELN
TypeBlog PostPublishedJan. 17, 2019, 1:38 p.m.... 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... -
CRISPR-mediated Plant Base Editors
TypeBlog PostPublishedJan. 3, 2019, 1:35 p.m.... 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 ... -
Rett Syndrome: A History of Research and Therapeutic Outlooks
TypeBlog PostPublishedNov. 19, 2020, 2:15 p.m....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... -
Plasmids 101: Broad Host Range Plasmids
TypeBlog PostPublishedApril 11, 2024, 1:15 p.m....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... -
CRISPR 101: Any Base Transversion Editors
TypeBlog PostPublishedApril 1, 2025, 1:15 p.m....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... -
AAV Titers: Where Do They Come From And What Do They Mean?
TypeBlog PostPublishedNov. 15, 2017, 3 p.m....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... -
An Inside Look into Addgene's Dev Team
TypeBlog PostPublishedJan. 23, 2020, 2:45 p.m....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 ...