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...p53) is one example of these difficult-to-study genes. Its main function is to regulate DNA repair, senescence...GC), TP53 was one of the most frequently mutated genes. Next, the authors explored how these variants of...ready to request from Addgene so you can combine the sensitivity of HiBiT with Addgene's quality control, ...luciferase enzyme (Figure 1). Addition of substrate generates a proportionate luminescent signal, strong enough...weak promoters, and even modest endogenous pools generate high signal-to-background luminescence (Schwinn...over time; and, importantly, repeated excitation generating reactive oxygen species that can lead to phototoxicity...Collection that express similar constructs available at Addgene. Read the original paper for more detail on whether...

...elegant bacterial immune system and an efficient gene editing tool… but boy does it have a lot of parts... resources on addgene.org  Addgene's CRISPR Guide Addgene's CRISPR 101 eBook  Addgene's CRISPR Cheat Sheet...section of the bacterial chromosome that stores a genetic record of previously encountered foreign nucleic...looser PAM requirements, which are available from Addgene!   RNA crRNA: “CRISPR RNAs” are transcribed from...CRISPR-Cas9 system for laboratory applications. At Addgene, we sometimes refer to the programmable guide sequence...sequence as the scaffold. gRNA: "Guide RNA" is a general term referring to the RNAs used by Cas proteins...Conclusion Ready for some more details? Check out the Addgene CRISPR guide or our CRISPR 101 ebook. We also ...

...line Y, or the phenotype of a mouse with a specific gene knock out, often you have to remember the experiment...Data Sharing Resources at Addgene.org Deposit Your Plasmids with Addgene Find Molecular Biology Protocols...innovation by all.  Additional Resources on the Addgene Blog Use Apps to Save Time and Track Your Experiments...

...this publication at Addgene. A protein-tagging system for signal amplification in gene expression and fluorescence...multiple protein molecules (and also available from Addgene!) Since the signal level provided by SunTag is ...applicable to studies of cell cycle progression, epigenetics and cellular reactions to external stimuli. It...pantheon of useful CRISPR/Cas9 techniques, we at Addgene are excited to see what’s coming next! References.... Find the plasmids from this publication at Addgene. Dynamic imaging of genomic loci in living human.... Find the plasmids from this publication at Addgene. ...

... GFP-containing organism,” says Hanson, “using a gene encoding one of the unexpectedly photoconvertible... Find plasmids from this publication at Addgene.   Addgene’s Fluorescent Protein Plasmids and Resources...multiple important technologies, including FRET and optogenetics. Even though GFP has been so thoroughly characterized... GFP is great news for microscopy enthusiasts! Addgene has empty backbones containing GFP variants used...PubMed. Find plasmids from this publication at Addgene. Subach OM, Patterson GH, Ting LM, Wang Y, Condeelis...

...approach is to target two exonic sites within a single gene using paired DSBs. This method allows for deletion...Medical School. Addgene is proud to present a video reprint of the CRISPR article "Generation of Genomic Deletions...within the deletion boundaries may be produced to generate clonal allelic series. Single DSBs resolved by...biologists can imagine, create, and examine novel genetic configurations. Watch the following video to learn...the full protocol from the JOVE article here on Addgene's website and find more CRISPR resources.    Thank... Medical School. His lab studies the molecular genetics of blood cell development and disease. Learn more...al. An erythroid enhancer of BCL11A subject to genetic variation determines fetal hemoglobin level. Science...

...they have been able to use CRISPR/Cas9 to generate mice with genes knocked out in the insulin signaling cascades...and Dr. Emmanuelle Charpentier. I was joined by Addgene Scientist Matt Ferenc (one of our resident CRISPR... to a year or greater with older technologies. Addgene is proud to be helping get CRISPR reagents out ...  New to CRISPR/Cas9 Technology? Learn More: Addgene's CRISPR/Cas9 Guide History of CRISPR Cas - A tale...

...CGE), which has been used in plasmid vaccine and gene therapy development to examine plasmid topologies...plasmid DNA concatemers in mammalian cells. J Gene Med. 2009 May;11(5):444-53. doi: 10.1002/jgm.1310. PMID: ...resources on Addgene.org  Addgene's Plasmids 101 eBookAddgene's Molecular Biology ReferenceAddgene's Restriction...length of the monomer (~11.6 kb). Image generated by SnapGene software.  Thanks to increasing affordability...RA, James AA, Kolodner R. recA-independent general genetic recombination of plasmids. Nature. 1981 Nov... performing restriction digests on plasmids at Addgene, we sometimes observe something odd in our uncut... the short-read Illumina MiSeq sequencing that Addgene uses.) However, when some researchers received ...

...specifically investigated protein binding at the TERT gene, but this method enables the unbiased discovery ...cDNA sequencing and RNA sequencing of complex heterogeneous RNA samples. Furthermore, such high processivity...

...Neddersen characterized a few samples by 16S rRNA gene sequencing which showed the presence of many “dark...Yellowstone National Park (YNP). Extremophiles - phylogenetically diverse microbes living in extreme physical...lab and ThermoCas9 from the van der Oost lab. Phylogenetic comparison indicates that like GeoCas9 and ThermoCas9...particularly useful. Additional resources on the Addgene blog Learn about other Cas proteins Read all of...GeoCas9, another thermostable Cas9 Resources on Addgene.org Read our CRISPR guide Find CRISPR plasmids for...

...Oregon created mTFP1 by transforming E. coli with a gene library of approximately 484 variants, including... your research Subscribe to the Addgene blog Resources on Addgene.org Learn about the Michael Davidson...Allele Biotechnology that now can be deposited at Addgene. What makes these fluorescent proteins unique and...was then screened through successive rounds of mutagenesis for the brightest teal fluorescent proteins that...scientists from Florida State University, they used a mutagenesis approach and found that specific amino acid changes...scientific community, please deposit the plasmids at Addgene! References Ai, Hui-wang, et al. "Directed evolution...PMCID: PMC3811051. Additional resources on the Addgene blog Read our fluorescent protein blog posts Find...

...during adeno-associated virus manufacturing." Human gene therapy21.10 (2010): 1251-1257. PMID: 20649475. ...have probably heard about Addgene’s viral service but did you know that Addgene also performs independent...101 eBook Additional resources on Addgene.org Learn about Addgene's viral service Learn about viral production...research? Since launching the viral service in 2016 Addgene’s research team has been actively investigating ...PMCID: PMC2957237. Additional resources of the Addgene blog Read our beginner's guide to viral vectors...

...Solubility Enhancer. In P. E. Vaillancourt, E. coli Gene Expression Protocols (Vol. 205, pp. 99–118). Humana...-59259-301-1:99 Resources on Addgene.org Addgene antibodies Addgene’s antibody guide Antibody protocols...having solubility issues!). Need a GST antibody? Addgene has you covered! His His tags, often called polyhistidine...Purification of His fusion proteins from E. coli is generally more straightforward and of a higher purity than...and other qualities of each tag class, they are generally best suited for their primary application. Ready...protocols  Resources on the Addgene blog Plasmids 101: Protein tags Antibodies 101: Epitope tags To Each...

...affect factors that could skew research results (gene expression, growth rates, etc.). Thus, it’s important...present in this line, making it genetically female. HEKs are generally cultured as an adherent monolayer...plasmids with the SV40 origin of replication. To generate a virus, you need only transfect the ‘helper’ .../fbioe.2021.796991 Additional resources on the Addgene blog Five Popular Model Organisms 10 Tips for...and Troubleshooting Additional resources on Addgene.org AAV Production in HEK-293T Cells  ...

...by the nearly infinite regulatory capabilities of gene pathways just waiting to be discovered. As we started...Bricking Science.      Additional Resources on the Addgene Blog Learn How You Can Use Video to Share Your... 10 Ways to Share Your Science Resources on Addgene.org Find Protocols and Protocol Videos Check out...

...at East Tennessee State University. He works with gene expression in Pseudomonas aeruginosa. Follow him...Reading, & Organizing Write for the Addgene Blog Resources on Addgene.org Check out Our Revamped Protocol...laborious, but helped build my understanding of how to generate a manuscript. At the end of the online course,.... Over time that portion of the paper will be generated. After you complete a publication quality figure...figure. Writing a scientific manuscript is like generating a quilt with your lab mates, you add the individual...100percentSDS.     Additional Resources on the Addgene Blog Dos & Don'ts When Publishing a Scientific...

...journal club discussion on Kevin Esvelt’s CRISPR/Cas9 gene drive paper was very popular. People kept asking... at Addgene.org Order Plasmids from Addgene! Find Free and Open Educational Resources at Addgene! ...BosLab. Once he optimizes plant cell culture and transgenesis he wants to share his knowledge with the community...microbes. We’ve ordered a few yeast plasmids from Addgene and are eager to do our first yeast transformations... @ZebraElement     Additional Resources on the Addgene Blog Experimenting in an Open Source Lab: from...

...regulation of DNA replication and gene transcription during vertebrate embryogenesis. My interests, though, have...Career Blog Posts Resources at Addgene.org Find Job Openings at Addgene ...than what fits for applied research. I focus in general terms on a couple of very broad topic domains, ...Pam_Hines and LinkedIn. Additional Resources on the Addgene Blog What Do I Do Now? Academic v. Non-Academic...

... to examine translation of Drosophila patterning gene oskar. Early in development, the osk-TRICK reporter...Find plasmids from this paper at Addgene. Additional Resources the Addgene Blog Learn More about SunTag... remained elusive. Two techniques developed by Addgene depositors have made it easier to track translation... on the nascent polypeptide recruit scFV-GFP, generating green puncta that colocalize with the red reporter...PMC2746483 Find ribosome profiling plasmids at Addgene. 2. Halstead, James M., et al. “Translation. An...: PMC4451088 Find plasmids from this paper at Addgene. 3. Yan, Xiaowei, et al. “Dynamics of Translation... Brightest Fluorescent Protein Resources on Addgene.org Find Fluorescent Protein Plasmids Check Out ...

... role in the transcriptional regulation of many genes. A number of groups have designed means of studying...with mAID. Using this method, the lab was able to generate conditional alleles of a few essential proteins... al., 2019). Find the mAID tagging toolkit at Addgene Yesbolatova et al., Methods, 2019. PubMed PMID:...targeted by the Cas9 nuclease. The Streptococcus pyogenes Cas9 (SpCas9) requires an NGG PAM to recognize...difficult. Find SpCas9 and Target-AID-NG plasmids at Addgene! Nishimasu H et al., Science, 2018. PubMed PMID... MerMAIDs - marine channelrhodopsins for the optogenetic toolbox Article contributed by Susanna Bachle...quickly and with high precision - a valuable optogenetics feature when studying neuronal activity. The...

...China was using CRISPR/Cas9 in an attempt to create gene-edited babies. Several hours later, the scientist...public consensus on genome editing, there was a generally agreed-upon international moratorium on utilizing...incorrectly known as “three-parent babies,” where fused genetic material from intended parents is “rehoused” in...decisions on scientific, ethical, regulatory, and general public consensus to the next meeting of experts...and the entire 2018 saga of “what’s happening in genetics this time?” She lives with her partner (in business...on theirpenmanship. Additional resources on the Addgene blog Find all of our CRISPR blog posts Download... genome engineering blog posts Resources on Addgene.org Read our CRISPR guide Find CRISPR plasmids by...

...Congratulations, you have a plasmid expressing your gene of interest (YGOI) and are ready to dive into your...received matches the expected sequence. Here at Addgene, we use NGS-based quality control to confirm the...necessary but also an easy process. Please vist Addgene's resource for plasmid verification to find additional...DNA gel.  More Plasmid Eductional Resources: Addgene's Molecular Biology Reference Find More Protocols...

...sequence between two target sites. Modifying multiple genes at once. Using multiplexed gRNAs can target multiple...simultaneous multiplex editing of six different genes. This system doesn’t rely on any additional or host-dependent...from this paper at Addgene. 9. Yin, Linlin, et al. “Multiplex Conditional Mutagenesis Using Transgenic ...plasmids from this paper at Addgene. Additional Resources on the Addgene Blog Browse Our CRISPR Topic...add multiple gRNAs on one plasmid. Sound good? Addgene has many tools to help you multiplex - we’ll use...multiplex gRNAs, some being: Using dual nickases to generate a knockout or edit. This can help reduce off-target...Multiplexing gRNAs: the basics One common question Addgene Senior Scientists receive is: can I express more...

...as a control. For easier portability, all viral genes are flanked by EcoRI and BamHI restriction sites...coronavirus or COVID-19 research, please email Addgene at help@addgene.org with your order number and so it can .... Sign up for email alerts on the Krogan lab's Addgene page to get notified!) Attacking coronavirus, one...reasons, the QCRG is delighted to be partnering with Addgene, who will be taking over the distribution to expand... coronavirus related plasmids and resources at Addgene* Many thanks to our guest bloggers Manon Eckhardt...

...Plasmids for Creating and Controlling Gene Networks Browse All SynBio Plasmids ... education policy. Additional Resources on the Addgene Blog Synthetic Biology and the Importance of Assembly...All Synthetic Biology Blog Posts Resources on Addgene.org Guide to SynBio Assembly Standards Find Plasmids...

...telephone, until the message reaches the nucleus where gene expression changes lead to cell proliferation and... on Addgene.org Find nanobody expression plasmids here Learn more about optogenetics in Addgene's optogenetics...induced binding. The lab used this general design scheme to generate two more OptoNBs, one against mCherry...loop 1 always generated OptoNBs with dark-induced binding, but insertion into loop 6 generated both blue ...on the key components of OptoNBs, how they are generated, and their use for protein purification and controlling...nanobody’s ability to bind its target protein.  Generating OptoNBs The opening and closing of the AsLOV2...spatiotemporal resolution than pharmacological or genetic manipulation of the signaling pathway.  As a proof-of-principle...

...that control cell cycle gene expression.  Additional Resources on the Addgene Blog Visit the fluorescent...work in Lin Tian’s lab at UC Davis is to generate a genetically encoded fluorescent sensor for serotonin...sequence an individual’s genome. But when they find genetic variants, what does that mean? Previous research...of Washington sought to address the effects of genetic variants in a high throughput manner. She helped...modeling, directed evolution, site-saturation mutagenesis, and medium-throughput screening, she increased...GFP Find microbiology blog posts Resources on Addgene.org Find fluorescent protein biosensors Browse Plasmids...

... observing low-copy genomic loci or single copy genes. If you want to visualize genomic loci brightly,...PP7, or boxB sequences inserted into the sgRNA generates blue, green, or red colors for labeling. A combination... PP7, or boxB sequences in the same sgRNA will generate additional colors. Image from Ma et al., 2016 ...CRISPR-Sirius a try. Find CRISPR-Sirius plasmids at Addgene. References Ma, Hanhui et al. “Multicolor CRISPR...PubMed PMID: 30377374. Additional resources on the Addgene blog Download our CRISPR eBook Learn about fluorescent...tools Read all CRISPR blog posts Resources on Addgene.org Browse our CRISPR plasmids for visualization...

...inhibiting gene expression, detecting neuronal signaling, or manipulating any other genetically controlled...wide array of genetic tools for neural interrogation like optogenetic tools, chemogenetic tools, and biosensors... the Addgene Blog Get Started with AAV Learn more about Lentivirus Find out More about Addgene's Viral...injected into the brain and efficiently deliver genetic information or tools, making them useful tracers...efficiently infects neurons, directs high-level transgene expression, and has retrograde functionality to...manipulated (e.g., activated or inhibited) by any genetic tool delivered by the AAV.   AAVrg requires high...evaluating how this serotype works in your system. Chemogenetic vectors  Expression of retrograde serotype vectors...

...Technique Q1: Is there any way to deliver a long gene (>4.5 kB) by AAV? A: One potential strategy is to...applications in diseases which may be linked to epigenetic changes, such as multiple sclerosis?  A: Basic...an exciting area for future AAV application to epigenetic editing. This area is discussed in our recent...excellent questions.    Additional Resources on the Addgene Blog Using virus in your research - a primer for...Tips for a first time AAV user Resources on Addgene.org Find Adenovirus Plasmids Adenovirus Guide Page...

...Suzuki et al. also envision eNLs as good reporter genes for endogenous protein fusions created using CRISPR...plasmids from this paper at Addgene. Additional Resources on the Addgene Blog Learn about Photoactivatable...proteins. To make luminescent imaging a reality, Addgene depositor Takeharu Nagai and colleagues at Osaka...nano-lanterns for use in various applications, including optogenetics, biosensors, and fusion proteins. Solving the...yield, and when transiently expressed in cells, generates much less signal than comparable fluorescent proteins... approach is especially suited to complement optogenetic tools, where light used for excitation can cause...cause unintended activation of the optogenetic system. Biosensors based on Nano-lantern technology are...

...at East Tennessee State University. He works with gene expression in Pseudomonas aeruginosa. Follow him...Protocols Check Out How Addgene Develops Lab Management Software Resources on Addgene.org Find Protoctols ... and do not necessarily represent the views of Addgene. I think it’s safe to assume that anyone reading...by New England BioLabs In my lab we do a lot of genetic construction and cloning experiments. Not all of...100percentSDS.     Additional Resources on the Addgene Blog Learn How to Keep a Lab Notebook for Bioinformatic...

...endonucleases. A given prokaryote typically has genes encoding one or a few restriction modification systems...Results Use REPLACR Mutagenesis to Mutate Your Plasmids Resources on Addgene.org Check Out Our Plasmid...newly synthesized product from a site-directed mutagenesis experiment? The answer to both questions is the...cleave DNA. DpnI is often used for site directed mutagenesis. During this process, incorporation of a desired...mutation into your plasmid of interest by PCR generates mutated plasmids with no methylation (there are...illustrates how sequence visualization software (we used Snapgene here) can help you assess whether your restriction...describing dam-/dcm- E. coli Resources on the Addgene Blog Keep up with the Latest Cloning Techniques...

...The first base editors revolutionized CRISPR gene editing. Cytosine base editors (CBEs) and adenine base...conducted a CRISPRi screen to identify endogenous genes that impact CGBE efficiency. In addition to using...review how CRISPR-derived base editing works in general. Base editors use either catalytically dead Cas9...efficiency and produced fewer indels than first-generation editors. This team also demonstrated that different...sequences) (Huang et al., 2024). BE4max, a CBE, generated similar deletions at a lower frequency. Their ...and its inclusion significantly reduced indels generated by CGBEs. What's next? Base transversion editing...base transition editing, we can expect several generations of optimization before the field begins to converge...

...In this case, Konermann et al. targeted MAPT, the gene encoding dementia-associated tau, delivering dCasRx...possibilities. To identify Cas13d, both groups used very general bioinformatic screens that looked for a CRISPR ...scientific sharing will continue to grow, and we at Addgene appreciate our depositors for making their tools...29551272 Plasmids from this paper are available at Addgene. Yan, Winston X., et al. “Cas13d Is a Compact ...29551514 Plasmids from this paper are available at Addgene. Learn more about research at Arbor Biotechnologies...Biotechnologies. Additional Resources on the Addgene Blog: Learn about CRISPR RNA Editing with REPAIR Get tips...Cas13a (C2c2) Characterization Resources on Addgene.org: Find RNA Knockdown Plasmids Check out our CRISPR...

...where we could incorporate cold-tolerance inducing genes within a regular mesophilic host system. This would...resources on the Addgene blog Find our synthetic biology blog posts Resources on Addgene.org Find plasmids... pC013-Twinstrep-SUMO-huLwCas13a plasmids from Addgene, which would potentially allow a stable Cas expression...” by Benoit Giquel, an outreach scientist from Addgene. The talk highlighted the importance of reagent...

...situ is the knock-in of fluorescent tags into the gene locus of interest. An advantage of this alternative...studied a synaptic protein that is linked to neurodegeneration. The level of this protein is decreased in...PMCID: PMC4121734. Additional Resources on the Addgene Blog Fluorescent microscopy techniques - Which... tagging strategies Additional Resources on Addgene.org Fluorescent protein plasmids Use FRET to study...protein-protein interactions Deposit plasmids with Addgene! ...

... widely available and applied by researchers for gene targeting. Fast forward a decade, and the CRISPR...Resources on Addgene.org CRISPR software and resources Addgene CRISPR Guide Addgene’s CRISPR 101 eBook... trans activity. AsCas12a, on the other hand, generates staggered DNA nicks at its target in cis and cleaves...end goal. CasPEDIA also features useful links to Addgene resources throughout the site. In the Experimental...each enzyme page, you'll find a link to search Addgene's collection for plasmids containing that enzyme... from the article if they're available through Addgene. So whether you're looking for the exact Cas9 plasmid... or browsing for the latest AsCas12a plasmids, Addgene and CasPEDIA have got you covered!     ...

...Chemical Modifications of CRISPR RNAs to improve Gene-Editing Activity and Specificity. JACS 144, 12584...resources on addgene.org  View Addgene's collection of empty gRNA vectors View Addgene's CRISPR resources...effectively serving their function of guiding Cas9 to generate a targeted break. These advances in gRNA technology...photocleavable gRNAs. Photocleavable guides are generated by introduction of a single photocleavable 2-nitrobenzyl...lentiviral plasmids Additional resources on the Addgene Blog  How to design your CRISPR gRNA CRISPR 101...

...repair of Cas9-generated breaks is useful if the intent is to make a knockout in your gene of interest, ..., as it is prone to generating indel errors. Indel errors generated in the course of repair by NHEJ are...are typically small (1-10 bp) but extremely heterogeneous. There is consequently about a two-thirds chance...Of some importance, the deletion can be less heterogeneous when constrained by sequence identities in flanking...overhangs is available. Cas9 can also be altered to generate a targeted single strand break; when two such ...  Dale Ramsden is a member of the curriculum in Genetics and Molecular Biology, the Dept. of Biochemistry... Break Diminishes Its Mutagenic Potential. PLoS Genet 5:e1000683 . https://doi.org/10.1371/journal.pgen...

...from Jiazhang Lian’s lab that enables simultaneous gene activation, repression, and editing with a single...studies.    Duong et al., Genetics 2021. https://doi.org/10.1093/genetics/iyab006       The CRISPR ... need. Here's what you'll find in this post: Epigenetic engineering with single-pot assembly Tracking...CRISPR plasmids New items from the viral service Epigenetic engineering with single-pot assembly By Eric ... of plasmids to the repository that will help epigenetic researchers custom design their own engineering...) Golden Gate assembly (backbones available at Addgene) with examples of potential synthetic chromatin...with BFP, sender cells expressing surface BFP generated BFP-labeled receiver cells. What’s more, G-baToN...

...downstream application such as quantitative PCR to assess gene expression. There are several things that researchers...everything, including your skin. Here are some general tips for working with RNA, even if you are using...10.1038/nprot.2006.83 Additional resources on the Addgene blog Read blog posts related to plasmid protocols...for purifying DNA without a kit Resources on Addgene.org Check out our molecular biology protocols Find...

...You realize, much too late, that you labeled your gene of interest incorrectly on slides 8-16. Your notes...remember related to this paper and/or biology in general until I reached Fig. 6, Panel L, and hope nobody...breath–you got this!  Additional resources on the Addgene blog Transferable Skills Guide: Time Management...

... useful as a mechanism of transient knockdown of gene expression. This system could be used in lieu of...complementary site in the genome where it will proceed to generate a blunt DNA double strand break. Cas9 has been...Cas12’s primary difference from Cas9 is that it generates staggered breaks, also known as sticky ends. Cas12a...targeting experiments requiring single strand DNA generation, such as homology directed-repair. Cas12a is ... of DNA. This makes it particularly useful in generating transient edits of signaling molecules (RNA) ...If your genome engineering goals expand beyond generating frameshift mutations or single base pair edits...s41586-021-03886-5 Additional resources on the Addgene Blog CRISPR 101: Which Cas9 do I choose for ...

...have been modified for genome editing, activating gene expression, visualizing genomic loci, and much more...plasmids from this publication at Addgene. Resources on the Addgene Blog Read about Cas9 Homolog Cpf1...effect on specificity was similar; SpCas9-HF1 generated fewer off-target cuts when compared to WT SpCas9...collaboration with the Joung Lab, they used targeted mutagenesis to create HypaCas9 (SpCas9 N692A/M694A/Q695A/...-HF1. The HypaCas9 plasmids are available from Addgene here! Future possibilities for Cas proteins With...26628643. Find plasmids from this publication at Addgene. 2. Nishimasu, Hiroshi, et al. "Crystal structure...26735016. Find plasmids from this publication at Addgene. 5. Chen, JS, et al. "Enhanced proofreading governs...

...cells)  that allow for continuous expression of genes integrated into the insect cell genome. Both of ...and cell free expression systems. Overall the general strategy for protein expression consists of transfecting...systems Yeast are a great expression system to generate large quantities of recombinant eukaryotic proteins... species due to its use as a model organism in genetics and biochemistry. When using S. cerevisiae, researchers...large-scale, stable production of proteins. In general yeast expression systems are easier and cheaper... use. Alyssa D. Cecchetelli is a Scientist at Addgene. She received her PhD from Northeastern University...proteins in plant root exudates Resources on the Addgene Blog Learn about inducible promoters Learn about...

...fewer transcripts of their antibiotic resistance genes and can therefore be cultured in media containing...plasmid prepping is a breeze. You get your stab from Addgene, streak for single colonies, sub-culture, and prep...use a different strain. For most applications, Addgene uses and recommends DH5𝝰! This strain consistently...density and plasmid yield as these factors are generally, but not perfectly, correlated. Typical growth...sufficient gas exchange in the overnight culture. Generally, large cultures in flasks may be shaken around... strain instructions from the manufacturer. On Addgene plasmid pages, you may see depositor recommendations...writing of this post. Additional Resources on the Addgene Blog Visit our Plasmids 101 Topic Page Learn How...

...error than normalizing to so-called housekeeping genes (Aldridge et al., 2008).  Woah! That was a lot of...quantification Normalizing to ubiquitously expressed genes Not as accurate; increased antibody costs; increased...Antibodies Resources on Addgene.org Addgene's Western Blot Protocol Addgene's Western Blot Protocol Video...Looking for a western blot protocol? Check out Addgene's western blot protocol and protocol video!  Antibodies...gel. Handcast gels are significantly cheaper and generate less waste. However, they also add time to an ...in size. However, these systems are expensive, generate plastic waste, and limit the amount of optimization... can only compare samples to a standard curve generated on the same blot. If your protein of interest ...

...to make sense of biology, with a special focus on gene editing techniques. References and resources References... versatile genome manipulation. Nature Reviews Genetics, 24(3), Article 3. https://doi.org/10.1038/s41576...DeepSpCas9, a deep learning–based model with high generalization performance. Science Advances, 5(11), eaax9249...). Chromatin context-dependent regulation and epigenetic manipulation of prime editing. Cell. https://...j.cell.2023.03.034 Resources More resources on the Addgene blog How to Design Your gRNA for CRISPR Genome ...and Flexibility to CRISPR Editing Resources on Addgene.org CRISPR: PRIME Edit Collection The CRISPR Guide...

...the expected complete sequence for the lacI/lacZ genes (1), while sequencing of white colonies showed a...PMCID: PMC6061786. Additional resources on the Addgene blog Learn the basics of Golden Gate assembly ...other plasmid cloning techniques Resources on Addgene.org Find basic molecular biology protocols Find ...