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We narrowed to 954 results for: App;

Showing: 701 - 750 of 954 results

  2. Cpf1 Update: Comparison to Cas9 and NgAgo

    Type
    Blog Post
    ...eclipse any of the CRISPR nucleases in AAV-based applications. Get More Cpf1 Background Here THE ORIGINAL ...the PAM sequence. Many research and clinical applications require HDR over NHEJ, so comparing HDR efficiency...

  3. Plasmids 101: FLEx Vectors

    Type
    Blog Post
    ...characterization of the location, morphology, and circuit mapping of a targeted neuronal population.    Figure... cell types for imaging and long-range circuit mapping. PubMed PMID: 18614669. PubMed Central PMCID: PMC2593125...

  5. Mobile-CRISPRi: Bringing CRISPRi to Diverse Bacteria

    Type
    Blog Post
    ... disease-associated. Mobile-CRISPRi: a modular approach for CRISPRi transfer Mobile-CRISPRi offers two...single-gene knockdown experiments. Mobile-CRISPRi applications Mobile-CRISPRi expands CRISPRi to non-model ...

  6. Production of Virus in Insect Versus Mammalian Cells

    Type
    Blog Post
    ...something you may have to optimize for your specific application. With mammalian systems, it is standard to do...titer the virus, or go straight into your viral application, like protein production. Titration of baculovirus...

  7. 10 Basic tips for mammalian cell culture

    Type
    Blog Post
    ...waste should be autoclaved before being disposed Happy culturing! Many thanks to our guest blogger, Sana...expression disregulation on HeLa cells." Journal of applied phycology27.6 (2015): 2321-2332. 3. Contamination...

  8. xCas9: Engineering a CRISPR Variant with PAM Flexibility

    Type
    Blog Post
    ... just trying to create a knockout, you might be happy targeting anywhere in an exon. But things get more...editing rates. This PAM flexibility should expand applications that require very specific targeting, like homology...

  9. Fluorescent Proteins 101: Fluorescent Protein Timers

    Type
    Blog Post
    ...possible consequences of FP aggregation, as Erik Snapp’s blog post shows. To avoid these issues, Subach ...-FT-N1 pTRE-Slow-FT pBAD/HisB-Slow-FT -  Applications of fluorescent protein timers FP timers have...

  10. Pooled Library Amplifications

    Type
    Blog Post
    ...to amplify the sample for further downstream applications. As individual libraries can vary significantly...feel free to contact us at [email protected]. Happy amplifying! More resources on the Addgene blog ...

  13. Light Sheet Fluorescence Microscopy

    Type
    Blog Post
    ...of LSFM microscopes (3,4). After this initial application, Stelzer’s group described the single-plane or...shadows along the illumination path. The shadows appear as stripes in the acquired image (5).  A way to...

  15. Which Fluorescent Protein Should I Use?

    Type
    Blog Post
    ...emission is green or red light. For example, T-Sapphire, LSSmOrange, and LSSmKate. Fluorescent Sensors.... al. (2010) “Fluorescent proteins and their applications in imaging living cells and tissues”  Physiol...

  16. Isolating Genomic Regions of Interest with the CRISPR System

    Type
    Blog Post
    ...and retroviral expression system for using this approach in in vivo studies (Fujita et al., 2018). In addition...DUX4, a transcriptional regulator that when inappropriately expressed can cause facioscapulohumeral muscular...

  17. Viral Vectors 101: The Retroviral Lifecycle

    Type
    Blog Post
    ...101: Types of Viruses Viral Vectors 101: Viral Applications Resources on addgene.org Addgene's Lentiviral...Liu, H., Xiao, H., Conway, J. A., Hunter, E., & Kappes, J. C. (1997). Functional RT and IN incorporated...

  20. Virus Protocol - Generating Stable Cell Lines

    Type
    Protocol
    ...expression observed using transient transfection approaches, generating cell lines using lentiviral vectors... tubes, VWR 21008-216 Lentivirus preparation Appropriate antibiotic for selection (e.g. puromycin, blasticidin...lower dilutions depending on your downstream applications. If you’ve titered your virus beforehand, you.... Add 1.5 mL of DMEM complete containing the appropriate antibiotic. This is the beginning of the selection...

  21. Affinity Purification of Recombinant Antibodies with Protein A or Protein G

    Type
    Protocol
    ...bottom. Add 5 mL of Pierce IgG Elution Buffer to the capped Gravitrap column and incubate for 10 min at RT...subsequent centrifugation step Pro-Tip Resin will appear compacted after centrifugation. Discard the flow...column in a new 50 mL LoBind collection tube. Slowly apply the sample to the center of the compact resin bed...exchange and concentration with an Amicon column Apply 15 mL of PBS to the reservoir of a Amicon Ultra-...

  22. Hot Plasmids - August 2020

    Type
    Blog Post
    ... have created a yeast-based directed evolution approach with some notable improvements. Directed evolution...specific guide RNA (gRNA) sequences. But these approaches rely on continual expression of both dCas9 and...

  25. Neuronal labeling with Spaghetti Monster

    Type
    Blog Post
    ...fluorescent proteins (FP) have also been important in mapping, monitoring, and manipulating neuronal circuits...combining advantages of FPs and epitope tags, smFPs appear to be very effective probes for finely visualising...

  26. Analyzing CRISPR Editing Results with ICE from Synthego

    Type
    Blog Post
    ...workflow - analyzing the data - is often under-appreciated or over-looked.  Synthego has developed a new...analysis Once the analysis is complete, a new screen appears with a graphical representation of the results ...

  28. Viral Production at Addgene

    Type
    Blog Post
    ...particles can be identified after negative staining and appear darker than full vector particles.     ...replaced with a new vial of early passage cells. Approximately two weeks post-thaw, cell culture supernatant...

  29. How to Be an Excellent Trainee

    Type
    Blog Post
    ...easiest to train individuals who are polite, appreciative, and clear about their needs. Remember, your...trainees one day when you are the senior personnel! Happy training! Resources Resources on the Addgene blog...

  30. How to Write a Scientific Review Article

    Type
    Blog Post
    ...papers and then feel ready to write. What actually happened was that each paper taught me a few things and...to read every single paper. I also learned to appreciate the discussion sections of the primary literature...

  31. Plasmids 101: Gateway Cloning

    Type
    Blog Post
    ...sequences into multiple vector systems. With the appropriate entry and destination vectors, one can use Gateway...elements to express the gene of interest in the appropriate system Recombines with the entry clone to generate...

  32. Antibiotic Resistance: An Old Solution but a New Problem

    Type
    Blog Post
    ...from random mutations in the genetic sequence that happen to be beneficial, and persist as a result. Why ...are susceptible to the drug, while the few that happen to possess resistance genes survive and multiply...

  34. CRISPR 101: Which Cas9 Do I Choose for My CRISPR Experiment?

    Type
    Blog Post
    ...Motif (PAM), the sequence NGG.  If these conditions apply to your experimental target, SpCas9 and its enhanced...specificity over a single SpCas9. The drawback of this approach is that two sgRNAs targeting opposite strands ...

  35. CRISPR Activation: A Practical Guide

    Type
    Blog Post
    ...suffer from an inability to easily deliver the appropriate isoform(s) of a given gene, as, many times, the...engineering parameters via a library-on-library approach. Nat Methods 12:823–826 . https://doi.org/10.1038...

  36. Plasmids 101: Control Plasmids

    Type
    Blog Post
    ...RNAi or CRISPR, non-targeting controls may be appropriate as they allow you to assess the specificity of...us now revisit our experiment. In Figure 1, it appeared as though the shRNA did not knock down expression...

  37. Plasmids 101: E. coli Strains for Protein Expression

    Type
    Blog Post
    ...plasmid-- make sure to maintain this by growing on appropriate media. Note: Purifying your expression plasmid...to make sure your cells have first reached an appropriate density. Cells in the exponential growth phase...

  38. Five Popular Model Organisms, Part 2

    Type
    Blog Post
    ...Escherichia coli as a Model Organism and Its Application in Biotechnology. In Escherichia coli - Recent...Physiology, Pathogenesis and Biotechnological Applications. InTech. https://doi.org/10.5772/67306 Koornneef...

  39. Plasmids 101: Expressing shRNAs for RNA Interference

    Type
    Blog Post
    ...world, there is a myriad of tools this could be applied to. When it comes to altering gene expression, ...is an umbrella term for a number of different applications that involve different types of small RNA molecules...

  40. Fluorescent Proteins 101: When GFP lets you down

    Type
    Blog Post
    ... proteins.Therefore, GFP is not suitable for applications that require anaerobic conditions. In experiments...protein module that can report on location. This approach enables the study of proteins in cells, but not...

  41. Deep Mutational Scanning with One Pot Saturation Mutagenesis

    Type
    Blog Post
    ...that expresses all desired variants of a protein. Applying a selective pressure winnows the pool down to ...technique described in Wrenbeck et al is a PCR-based approach for generating a customizable comprehensive mutagenesis...

  42. Transferable Skills Guide: Time Management

    Type
    Blog Post
    ... cell splitting issue at all if I’d planned appropriately. Back in my undergrad days, I had general goals... do I write things down? It’s more viscerally appealing for me to physically cross things out (it just...

  43. Antibodies 101: Introduction to Gating in Flow Cytometry

    Type
    Blog Post
    ...adjust this setting as needed. To figure out the appropriate voltages to observe your cells on a flow plot...voltage describes an electric potential that can be applied to the photomultiplier inside the cytometer to ...

  44. How to Negotiate a Successful Lab Start Up Budget

    Type
    Blog Post
    ...discounts, collaborations, access to internal award applications, etc. If this is the institute that can help...budgets and invite you to leave a comment below Happy Sciencing, Damien   Damien Wilpitz is a laboratory...

  45. CRISPR Library Amplification

    Type
    Protocol
    ...sufficient quantities of library for experimental applications. Repeated amplifications should be avoided as... of a library of 100,000 plasmids. Colonies may appear small and require extra incubation time in order... as the quantification method. If all Maxipreps appear to contain sufficient DNA for use, pool samples...purifications. Consult the manufacturers handbooks for appropriate volumes and numbers of tandem purifications. ...

  46. Plasmids 101: Dimers and Multimers

    Type
    Blog Post
    ...something odd in our uncut DNA control: a band or two appear on an agarose gel at notably higher molecular weights... only in the uncut DNA; the rest of the digest appears normal and produces the expected fragments for ...

  47. Protocol - How to Streak a Plate

    Type
    Protocol
    ...(with appropriate antibiotic) Bacterial stab Procedure Obtain an LB agar plate with appropriate antibiotic...working near a flame or bunsen burner. Obtain the approrpriate bacterial stab or glycerol stock . Using a sterile...

  48. Cancer and the Immune System: Deciphering the Relationship

    Type
    Blog Post
    ...immunotherapeutic agents. Currently the FDA has 12 approved monoclonal antibodies targeting the PD-L1 pathway...Dendreon Corporation is currently the only FDA-approved DC therapy for prostate cancer and is aimed at...

  49. Protocol - How to Inoculate a Bacterial Culture

    Type
    Protocol
    ...add liquid LB to a tube or flask and add the appropriate antibiotic to the correct concentration ( see...Large plasmids usually have a low copy number (approximately one or two copies per cell) and they need to...to grow for longer periods of time (approximately 18-30 hr). On the other hand, smaller plasmids can be...

  50. Protocols for Molecular Biology, Plasmid Cloning, and Viral Preps

    Type
    Protocol
    ...protocols that you can use for a wide range of applications, with videos for select protocols in the right-hand...preparations. Antibodies Protocols for common antibody applications. Intro to the Lab Bench Name Description (Link... of DNA to a plasmid Gibson Assembly Combine overlapping DNA fragments in a single reaction Ligation Independent...
Showing: 701 - 750 of 954 results