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We narrowed to 672 results for: abo.1

Showing: 621 - 640 of 672 results

  1. Bacterial Toxin-antitoxin Systems as Molecular Biology Tools

    Type
    Blog Post
    ...plasmids are passed down through generations (Figure 1). These so called “plasmid addiction systems” require... more about CcdB, the toxic key to efficient cloning Read all Plasmids 101 blog posts Read about knockout...with the other toxin-antitoxin systems described above, one part of the pair is more stable than the other...

  2. Viral Production at Addgene

    Type
    Blog Post
    ...determine purity of the AAV preparation. Figure 1: Silver staining of purified and non-purified AAV ... before being distributed to customers. Details about our production protocols, titering methods, and ...either in-house by Addgene scientists or through collaboration with viral vector manufacturing facilities, ...generated from lentiviral preparations (described above) that are subject to a concentration step prior ...

  3. Plasmids 101: Choosing an Antibiotic Resistance Gene

    Type
    Blog Post
    ...antibiotic selection. Essentially, this gene is a 2-in-1 providing both bacterial and eukaryotic selection,...therefore prevent cell death. One of the cool things about amp is that it's exclusively toxic to dividing cells...antibiotic selection. If you feel like multiple of the above antibiotics would work for you, they likely do! ...

  4. Plasmids 101: Shuttle Vectors

    Type
    Blog Post
    ...designed to shuttle between species. Figure 1: Shuttle vectors are vectors that can function in ...your experiment, meaning you don’t have to worry about vector replication at all. In these cases, transient...place in. In many cases, you won’t need to worry about the expression of your gene of interest in both ...

  6. New CRISPR Tools: Cas7-11 and PASTE

    Type
    Blog Post
    ... of low-toxicity RNA editing tools.   Fig. 1 Schematic of luciferase RNA knockdown assays using.... Eugene Koonin and Kira Makarova, longtime collaborators of Omar and Jonathan and experts in CRISPR evolution...

  7. Viral Vectors 101: Preparing Pooled Libraries

    Type
    Blog Post
    ...your virus. Let’s start there!     Figure 1: Flowchart showing the process for using pooled lentiviral...ordering a pre-made library, you don’t need to worry about amplifying and producing lentivirus. But pre-made...

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

    Type
    Protocol
    ...Choose Option 1 or Option 2 based on the concentration of the pooled sample above. Option 1: Buffer exchange...NaH 2 PO 4 ∙H 2 O 1 L deionized water Adjust pH to 7.0 Autoclave or filter sterilize 1 M sodium phosphate...NaH 2 PO 4 ∙H 2 O 1 L deionized water Adjust pH to 7.0 Autoclave or filter sterilize 1 M of sodium phosphate...antibody to 1 mg/mL with PBS if needed. For long term storage, add sterile sodium azide to 1 mM. Option...start with about 250 mL of supernatant and add 250 µL of 100X protease inhibitor cocktail. Add 1 part Protein...concentration of the pooled sample is above 1.0 mg/mL proceed to Option 1 with a buffer exchange using a Zeba...purify recombinant antibodies. Workflow Timeline Day 1: Purify antibody Day 2 or later: Buffer exchange Equipment...

  9. Protocol - pLKO.1 – TRC Cloning Vector

    Type
    Protocol
    ...of Contents A. pLKO.1-TRC Cloning Vector A.1 The RNAi Consortium A.2 Map of pLKO.1 A.3 Related plasmids...Order oligos compatible with pLKO.1 C. Cloning shRNA oligos into pLKO.1 C.1 Recommended materials C.2 Annealing... References H.1 Published articles H.2 Web resources I. Appendix I.1 Sequence of pLKO.1 TRC-Cloning Vector...information Back to Top A. pLKO.1-TRC Cloning Vector A.1 The RNAi Consortium The pLKO.1 cloning vector is the backbone...marker encoded in pLKO.1 allows for convenient stable selection. Figure 1 : Map of pLKO.1 containing an shRNA...puromycin should be from 1-10 μg/mL in 1 μg/mL increments. d. Label plates from 1-10 and add appropriate...Appendix I.1. Sequence of pLKO.1 TRC-Cloning Vector Click here to see the sequence of pLKO.1 TRC-cloning...

  10. Lab Safety for Biosafety Levels One and Two

    Type
    Protocol
    ... provides information for both biosafety level 1 (BSL-1) and biosafety level 2 (BSL-2). The purpose of... BSL-2 Guidelines Remember, the BSL-1 laboratory guidelines above are expected to be followed in addition...guidelines provide steps to ensure you are working in BSL-1 and BSL-2 labs safely. Protocols...Safety for Biosafety Levels One and Two (BSL-1 and BSL-2) Intro to the Lab Bench Check out more protocols...each level has different safety requirements. BSL-1 is designated for those working with microbes that...BSL-2 includes all of the precautions needed in BSL-1, along with additional precautions to prevent injuries...container Fire blanket Fire extinguisher Guidelines BSL-1 Guidelines Before You Work Right after entering the...

  11. Using Video to Share Your Science: We Share Science

    Type
    Blog Post
    ...email Ryan at [email protected]   References 1. Larivière, Vincent, Yves Gingras, and Éric Archambault...learn about how researchers are using “big data” to look at Lyme Disease in new ways. Or learn about the...ot­her disciplines, are even less likely to read about our work. We therefore must discover new ways to.... Only on occasion do we then have time to read about research from other disciplines, or from other countries...communication through video     Video abstracts about research, especially published research, have grown...researchers have Facebook pages, few share any details about their research itself. Pinterest is also a favorite...created a new platform for sharing video abstracts about research: www.WeShareScience.org. WeShareScience...

  12. 5 Reasons to Use Reddit for Science Communication

    Type
    Blog Post
    ...first foray into Reddit. Here are some reasons why: 1. Reddit has a large and active community of people...not too worried about selling yourself and really do just want to get information about yourself and your...where users can participate in forum discussions about a wide variety of topics. Individual forums are ...where users can ask designated scientists anything about a particular topic. Given Joanne’s expertise in... is well moderated. One of our original worries about sharing science advice on Reddit was that we’d be...answering questions. r/Science has pretty strict rules about not being too sales-y in posts which falls in line...any scientists to refer to if they have questions about science careers, women in STEM, work-life negotiation...

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

    Type
    Blog Post
    ...conference for any reason was a Singleton. Figure 1: The Binning Singletons logo. Image: James et al.,...noticed Holly Bik live-tweeting about topics I needed to know more about. I caught up with her outside ...me for coffee and spent an hour with me talking about the tools I could use, and some experimental hurdles...other folks at the meeting. I didn’t know anything about these people at the time, but I met some very accomplished... my mind because we’d had discussions in my lab about whether or not to keep DNA sequencing singletons...singletons out, and there is healthy debate online about this (our lab keeps singletons). I felt like anyone...scientific conferences Read our ECR Toolbox blog post about using social media as a scientist ...

  14. Filming Science Videos in the Age of Social Distancing

    Type
    Blog Post
    ... over again. Here’s that process!   Figure 1: The Design-Build-Test-Learn cycle adapted for video... Production Specialist, I’m constantly thinking about ways to improve our productions to deliver the best...four months after I started with Addgene, I was about to have a check-in with my boss Jennifer. I had ... would be a couple months. Jennifer and I spent about an hour that morning scrambling. Everything we had...someone to be comfortable on camera while thinking about things like lighting, camera angle, and shot composition... allowing people to sit without having to worry about getting in close to mic them up. Monitor: I then...first video in our Intro to the Lab Bench series about PPE! We made this with a combination of in-lab filming...

  15. Transferable Skills Guide: Creativity

    Type
    Blog Post
    ...yourself in a creative rut, read on!   Fig 1: Blugene using creative problem-solving in the lab...brainstorming sessions.  Allow the group time to think about the problem before you meet. This holds the dual... meetings run smoother when people have thought about the issue ahead of time. Encourage dissent. The ... decisions while encouraging the group to think about alternative options. When a devil’s advocate is ...a few minutes every month dedicated to thinking about the future will improve your ability to innovate...to be able to continue our experiments. Thinking about these plans before you’ve run out of consumables...

  16. Overcoming the AAV Size Limitation for CRISPR Delivery

    Type
    Blog Post
    ...indel formation and phenotypic changes as early as 1 week post-injection. Livers from these mice were histologically...Addgene Blog Learn more about Cas9 Variants and the Expanded CRISPR Toolbox Read about CRISPR enzyme Cpf1 ... and specific? The two AAV strategies described above showed successful target modification, indicating...

  17. Plasmids 101: Stringent Regulation of Replication

    Type
    Blog Post
    ...levels of DnaA to get the job done.   Figure 1: DNA melting by DnaA and IHF is the first step of ... negative regulators. The genome of E. coli is about 4.6 million base pairs long and contains a single...copy number and compensating for the additional metabolic cost imposed on the host with a beneficial function.... On the other hand, they also impose a lower metabolic cost on cells and produce lower amounts of protein...

  18. AAV Production in HEK293 Cells

    Type
    Protocol
    ...7.5% Sodium Bicarbonate, 7.5 mL 1 M HEPES to 750 mL DMEM + 1 g/L glucose. 1 mg/mL polyethylenimine (PEI) ...Calculate the amount of each plasmid needed to have a 1:1:1 molar ratio with 2 mg total DNA per CS5 Plasmid ... determine the total μg/bp we need to achieve a 1:1:1 molar ratio of each plasmid: 2000 μg / 24,961 bp...deionized water + 100 mL of 1 M Tris HCl pH 8.5 + 60 mL of 5 M Sodium Chloride + 4 mL of 1 M Magnesium Chloride...high glucose, Corning 10-013-CV DMEM, low glucose (1 g/L) glucose, sodium pyruvate, Corning 10-014-CV (... sodium bicarbonate, Corning 25-035-CI (optional) 1 M HEPES, HyClone SH30237.01 (optional) L-alanyl-L-glutamine...cations can affect the attachment of adherent cells) 1 mg/mL Polyethylenimine (PEI) 25 kDa MW Pro-Tip Other...

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

    Type
    Blog Post
    ...complex. So how exactly does this work? Figure 1: APOBEC- and Cas9-mediated editing (ACE) reporter ...for base-editing activity The reporters mentioned above all rely on GFP fluorescence and cell sorting using... Additional resources on the Addgene blog Read about other CRISPR reporters Browse our CRISPR blog posts...

  20. Kit Free RNA Extraction

    Type
    Protocol
    ...tissues: use 1 mL of Solution D per 100 mg of cells. For cultured cells: use 1 mL of Solution D per 1 X 10 7...tissues: use 1 mL of TRIzol® per 100 mg of cells. For cultured cells: use 1 mL of TRIzol® per 1 X 10 7 cells... to 1 mL of lysate: Add 0.1 mL of 2 M sodium acetate (pH 4.0), mix thoroughly by inversion. Add 1 mL water-saturated...Option A): Add 1 volume of Isopropanol to the extracted aqueous layer. Incubate at -20°C for 1 hour. Lithium.... Add 0.2 mL of Chloroform/Isoamyl alcohol (49:1) per 1 mL of TRIzol® used. Shake vigorously by hand for...with the volatile reagents in the list above. Procedure Option #1 - Solution D Protocol Before starting...
Showing: 621 - 640 of 672 results