Skip to main content
Addgene

Enter a sequence to perform a BLAST-based search of Addgene plasmid sequence databases.

In Category

We narrowed to 1,019 results for: CAL;

Showing: 721 - 740 of 1019 results

  1. Reaching out to China: Canton Nucleic Acids Forum (CNAF) 2015

    Type
    Blog Post
    ...remarkable progress on establishing a system for the chemical replication of RNA in the absence of evolved enzymatic... about using CRISPR/cas to address the growing clinical problem of antibiotic resistant bacteria. See ... and Sun Yat-sen University presented two new technical developments. Using the Tet-on system, his lab...

  2. xCas9: Engineering a CRISPR Variant with PAM Flexibility

    Type
    Blog Post
    ...edits or knock-ins. Editing efficiency decreases drastically once your cut site is more than 10 bp away from... mutagenesis plasmid (MP6) permits quick, large-scale mutation of the gene of interest. They created a... a selection phage (SP) to carry catalytically dead SpCas9 (dCas9) fused to the bacterial polymerase subunit...

  3. Multiplexed Capture of Promoter-enhancer 3D Chromatin Structures Using CRISPR

    Type
    Blog Post
    ...but close together in terms of where they are physically located. DNA is then sheared into pieces small...on-target enrichment. 2. Determining the spatial and hierarchical organization of chromatin interaction networks...CAPTURE 2.0 is the construction of spatial and hierarchical chromatin interaction networks. Think of these...

  4. CrispyCrunch: High-throughput Design and Analysis of CRISPR+HDR Experiments

    Type
    Blog Post
    ...sequencing primers for quality control. At such a scale, automation is crucial, both for speeding things...to the same principles listed here. It employs biological best practices. For example, it ranks guides ...published on how to do it best. Scientists are typically advised to mutate the PAM to which the guide binds...

  5. Plasmids 101: Multicistronic Vectors

    Type
    Blog Post
    ...bicistronic vectors take advantage of an element called an Internal Ribosome Entry Site (IRES) to allow...use the 2A peptides instead of an IRES when stoichiometrically equivalent levels of expression are required...journal.pone.0018556. Epub 2011 Apr 29.) PubMed. 5. Scalable signaling mediated by T cell antigen receptor-...

  6. An Introduction to Adenovirus

    Type
    Blog Post
    ...culprit for these unusual cases was either 1) a physiological effect of SARS-CoV-2 toward destabilizing one...Adenovirus (and/or to its partner, AAV) or 2) a sociological effect of the COVID-19 lockdowns causing a lack...Adenoviruses: General Features. In Reference Module in Biomedical Sciences. Elsevier. https://doi.org/10.1016/B978...

  7. CRISPR 101: Making a Knock-In Cell Line

    Type
    Blog Post
    ...not the case for knock-ins. Knock-in mutations typically require an exact DNA sequence to be incorporated...know that HR is often mutated in cancer cells, specifically in breast and ovarian lines commonly used in...common editing pathway for DNA repair. To tilt the scales in your favor for an HDR edit, you can try one ...

  8. Simplify Cloning with in vivo Assembly

    Type
    Blog Post
    ...-55 °C for the homologous region (calculated using Oligo Calculator). Insertions - To insert short sequences...parental DNA (37 °C for 15 mins). This enzyme specifically cuts methylated DNA, so it will selectively ...

  9. CRISPR 101: Ribonucleoprotein (RNP) Delivery

    Type
    Blog Post
    ...Liang et al. 2017).  PEG-mediated methods, while technically less difficult to perform, often show low efficiency...ribonucleoprotein electroporation of zygotes." Journal of Biological Chemistry (2016): jbc-M116. PubMed PMID: 27151215...Cell-Type-Specific Gene Editing." Journal of the American Chemical Society 140.21 (2018): 6596-6603. PubMed PMID:...

  10. DNA Purification Without a Kit

    Type
    Blog Post
    ...methods avoid many of the drawbacks of kits and typically follow the same principles: cells are lysed, RNAs...method described in Kim and Morrison is nearly identical to that of commercially available kits but instead...B., & Gillespie, D.A. (1979). Preparative and analytical purification of DNA from agarose. Proceedings...

  11. The Importance of a Fun Workplace: Company Culture at Addgene

    Type
    Blog Post
    ... familial aspect of Addgene. As our size has drastically increased, it would have been a tight squeeze...event! Addgene company culture At Addgene we organically developed the philosophy of  “work hard, play...department) right in your breakroom. 5. Throw some physical activity into the mix It’s easy to sit around ...

  12. Negotiating Work and Life: How to Find the Joy

    Type
    Blog Post
    ...community to help Involve family. If you don’t have local family to help, create “fake” family by forming ...help you do your job better. #8 Thou shalt enthusiastically foster independence Your partner can make ... fold their laundry, take public transportation, call a cab and remember their own school assignments,...

  13. Tips for Writing a Good Cover Letter

    Type
    Blog Post
    ...writing are key skills. Other fields, often highly technical or skills-based ones, may not use them very often...questions about your application. These could be logistical, like briefly explaining a gap in your job history...letter needs to be coherent and free of typos and grammatical errors. It does not need to be a literary masterpiece...

  14. Viral Production at Addgene

    Type
    Blog Post
    ... relative to empty particles (black arrowhead). Scale bar = 100 nm.   AAV resources Browse AAV preps... a concentration step prior to being frozen. Specifically, lentiviral particles are collected from the... strain of E. coli. In addition, plasmids are typically prepared using endotoxin-free plasmid purification...

  15. Save Time with Transient Plant Leaf Transformations

    Type
    Blog Post
    ...below. The GUS gene (β-glucuronidase) with histochemical GUS-staining is a very useful visual reporter...protein of interest to elucidate the subcellular localization of your protein of interest. Find all plasmids...see Figure 3). This method is probably the most scalable method. Transfer your plant into the dark 16 ...

  16. Four Base Editing Reporters to Monitor and Enrich Editing in Real-time

    Type
    Blog Post
    ...are subject to excision and ssDNA cleavage by canonical base excision repair enzymes. At the same time...targeted by a cytidine deaminase base DNA editor. Specifically the BFP mutant (BFPH66) contains a histidine...markers, and other enzymes such as Cre-recombinase drastically expanding the base-editing reporter toolbox. ...

  17. Binning Singletons: Tackling Conference Networking When You Don’t Know Anyone

    Type
    Blog Post
    ...by Joe James from Binning Singletons. The sheer scale of a large conference can be intimidating. And it...when we bring all the bins together at a mixer. We call this Quorum Sensing, after the phenomenon in bacteria... career scientists, students, and those from historically underrepresented groups. Through such mentoring...

  18. Overcoming the AAV Size Limitation for CRISPR Delivery

    Type
    Blog Post
    ...SpCas9 and SaCas9, Zhang’s group used an approach called BLESS (direct in situ breaks labeling, enrichment...post-injection. Livers from these mice were histologically normal and liver injury markers were not increased...enough to deliver with AAV. This system fuses catalytically dead dCas13b to the catalytic domain of RNA ...

  19. The PAM Requirement and Expanding CRISPR Beyond SpCas9

    Type
    Blog Post
    ... D1135E variant is far more selective for the canonical S. pyogenes PAM sequence NGG compared to wild-...novel CRISPR proteins has and will continue to dramatically increase the number of CRISPR applications. ...Wolfe SA, Sontheimer EJ (2018) NmeCas9 is an intrinsically high-fidelity genome-editing platform. Genome...

  20. Antibodies 101: Reading a Flow Plot

    Type
    Blog Post
    ...generate a dot plot (Figure 1). Clustered events are called a population.                             ...of markers you use to define your population is called your “gating strategy.” This series of markers ...around clusters to define populations. This shape is called a ‘gate’; it tells the machine that everything ...
Showing: 721 - 740 of 1019 results