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  1. Developing a cold tolerant E. coli using specialized chaperones

    Type
    Blog Post
    Published
    Jan. 28, 2020, 2:35 p.m.
    ...inefficiently that no industrial operation would ever consider them worth the trouble.  Enter ELSA: E. coli Learning...for research.  We’ve also involved ourselves in extensive outreach programs to encourage and embolden the...

  2. Antibodies 101: ChIP

    Type
    Blog Post
    Published
    March 10, 2022, 3:25 p.m.
    ...is an extremely useful technique that provides insight into protein:DNA interactions. ChIP works by using...exploring these interactions, and can give valuable insight into DNA expression and protein interactions, including...

  3. Viral Vectors 101: Viral Applications

    Type
    Blog Post
    Published
    April 13, 2023, 1:15 p.m.
    ...integrated into the genome of the target cell or transiently expressed. It’s important to note that for all...vector can express your construct in target cells transiently or constitutively depending on the virus used...

  4. Plasmids 101: Sequence and Ligation Independent Cloning (SLIC)

    Type
    Blog Post
    Published
    Sept. 26, 2024, 1:15 p.m.
    ...; or by incomplete PCR, which skips the final extension step to retain products with single-stranded overhangs...assembly is cost, as T4 polymerase is much less expensive than the enzymes required for Gibson assembly....

  5. CasPEDIA: A Functional Classification of Cas Enzymes

    Type
    Blog Post
    Published
    Jan. 25, 2024, 2:15 p.m.
    ...Cas enzymes and variants available. Without a comprehensive resource, it can be overwhelming to choose a...resources throughout the site. In the Experimental Considerations section on each enzyme page, you'll find a ...

  7. Plasmids 101: Methylation and Restriction Enzymes

    Type
    Blog Post
    Published
    June 30, 2016, 2:30 p.m.
    ... and gotten unusual or unexpected results? Or considered why DpnI will degrade your template DNA from ...level, the potential for methylation should be considered when digesting DNA. Why? Well, even though Dam...

  9. Lighting Up Cell Signaling with Photoswitchable Kinases

    Type
    Blog Post
    Published
    July 25, 2017, 1:20 p.m.
    ...phosphorylated MEK1/2 levels, suggesting PP1/PP2A is responsible for this MEK1/2 negative feedback. 3. Studying...Silvia Carrasco, and Markus W. Covert. "High-Sensitivity Measurements of Multiple Kinase Activities in...

  10. Choosing the B(right)est Fluorescent Protein: Aggregation Tendency

    Type
    Blog Post
    Published
    June 15, 2017, 2:30 p.m.
    ...be used to measure your fluorescent protein’s propensity to aggregate. The basics and pitfalls of these... the best way to test a fluorescent protein’s propensity to homodimerize. Several studies have used the...

  11. Plasmids 101: CcdB - The Toxic Key to Efficient Cloning

    Type
    Blog Post
    Published
    Dec. 8, 2016, 3:30 p.m.
    ...toxin-antitoxin system encoded by the ccd operon, which is responsible for plasmid maintenance during cell division....E. coli do not contain the F plasmid (and are considered F-); however, there are a few popular lab strains...

  13. Seven Tips for Using LinkedIn as a Scientist

    Type
    Blog Post
    Published
    Oct. 1, 2019, 1:21 p.m.
    ...your network. If someone has connected with me, I consider them fair game for connecting to others.  Pro ...including and especially the hiring manager. This is considered intrusive and it’s still connecting with a stranger...

  14. Troubleshooting Your Plasmid Cloning Experiment

    Type
    Blog Post
    Published
    Sept. 24, 2019, 12:55 p.m.
    ...Streak on them an E. coli  strain known to be sensitive to that antibiotic, and if the strain is growing...positive clone after these steps, you might want to reconsider your approach, and try a different cloning strategy...

  15. The Breast Cancer Microenvironment: A Tumor’s Backstage Team

    Type
    Blog Post
    Published
    Jan. 31, 2019, 1:32 p.m.
    ... transport miRNA from drug-resistant cells to sensitive ones. Though this process is not fully understood...malignancy, concluding that these cells may be considered "natural tumor suppressors." A growing interest...

  16. What's New in CRISPR - November 2021

    Type
    Blog Post
    Published
    Nov. 18, 2021, 2:15 p.m.
    ...increase activity and decrease the gRNA size by intensively remodelling the gRNA scaffold through five rounds...tools, the desired edits are templated by the 3′ extension of the pegRNAs. PEDAR is capable of correcting...

  18. New Tool for Lineage Tracing: The ClonTracer Library

    Type
    Blog Post
    Published
    Sept. 22, 2015, 2:30 p.m.
    ...abundance before and after a treatment. The library consists of a pool of lentiviral vectors with roughly 73...populations, they found that similar barcodes consistently made up the majority of the resistant populations...

  19. 6 Tips for Grant Writing

    Type
    Blog Post
    Published
    Oct. 18, 2016, 2:30 p.m.
    ...in different fields that can provide different insight into an area - not just working within the established...collaborators can also help you get some critical insight into what you're doing wrong. All this helps to...

  20. Popular Retroviral Vectors and Their Uses in Scientific Research

    Type
    Blog Post
    Published
    July 24, 2018, 12:51 p.m.
    ... 1000000097) The Winslow lab barcoding library consists of 96 retroviral plasmids (based on the Murine...mammalian cells. The MuLE system kit from the Frew lab consists of 91 building block Entry vectors and 5 lentiviral...

  22. Adeno Associated Virus (AAV) for Cell and Gene Therapy

    Type
    Blog Post
    Published
    Nov. 7, 2017, 1:59 p.m.
    ...lack all viral genes, including those that are responsible for integration into host chromosomes, further...to other gene delivery tools that are under consideration, recent advancements in various AAV technologies...

  23. Nanoblades: Tiny CRISPR Ninjas for Genome Editing Difficult Cells

    Type
    Blog Post
    Published
    Sept. 26, 2019, 12:50 p.m.
    ...and mice with Nanoblades CRISPR has also been extensively used to create transgenic mice by microinjecting...post Resources on Addgene.org Check out our comprehensive CRISPR guide Find CRISPR plasmids and resources...

  24. Mouse Modeling, Part 2: Breeding and Crossing Mice

    Type
    Blog Post
    Published
    Aug. 6, 2019, 12:55 p.m.
    ...Special breeding cases Whether your mouse colony consists of genetically engineered mice with a single mutation...strains that have been crossed, there are many considerations for effectively breeding your mice. Because...

  28. Plasmids 101: In-house NGS Plasmid Quality Control Now at Addgene

    Type
    Blog Post
    Published
    April 6, 2021, 12:30 p.m.
    ... example, the CAG promoter, a hybrid promoter consisting of a CMV enhancer, chicken beta actin promoter...any template DNA and primers.  After performing extensive QC on that sample, we’ll then load it on to our...

  29. Viral Vectors 101: Chemogenetics

    Type
    Blog Post
    Published
    July 11, 2023, 1:15 p.m.
    ... crossing the blood-brain barrier.  Another consideration when picking ligands for chemogenetic studies...response you aim to manipulate. It is important to consider that the same receptor can elicit different responses...

  30. Antibodies 101: Stripping and Reprobing Western Blots

    Type
    Blog Post
    Published
    Aug. 1, 2024, 1:15 p.m.
    ...abundance of your targets isn’t too different, consider antibody affinity: stronger binding antibodies...membranes recommend re-wetting with methanol before rinsing with wash buffer, as dry PVDF won’t evenly re-absorb...

  31. Progress Towards a PAM-Free CRISPR

    Type
    Blog Post
    Published
    March 7, 2024, 2:39 p.m.
    ...the ‘quality’ of the gRNA sequence, including propensity for off-targets, secondary structures, etc., ...more pan-genome PAM flexibility, you’ll want to consider one of several Cas9s that recognize NG PAMs. The...

  32. Corralling and Cornering the Goldilocks Antibody

    Type
    Blog Post
    Published
    Aug. 9, 2022, 1:15 p.m.
    ...scientists to confidently choose reliable and consistent antibodies for specific markers. The Screening...production methods    NeuroMab and others have an extensive library of monoclonal antibodies; Addgene collaborates...

  33. Engaging with science and society at pgEd

    Type
    Blog Post
    Published
    Sept. 20, 2018, 12:17 p.m.
    ...institutions. The conversations often move quickly into sensitive topics and uncomfortable places, from sticky ethical...rapidly changing landscape of genetics and our consistent stream of events mean that there is never a dull...

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

    Type
    Blog Post
    Published
    Dec. 1, 2015, 3:30 p.m.
    ...using bacterial phage vectors with CRISPR/cas9 sensitizing gene cassettes. Thse vectors are engineered to...genes in the bacteria thus making the bacteria sensitive to known antibotics. These might be used on hospital...

  36. Story of a SynBio Startup: RevBio's Epiphany (or Lack Thereof)

    Type
    Blog Post
    Published
    June 5, 2014, 3:32 p.m.
    ... a company. Revolution Bioengineering is two responsible adults in their 30s who have quit their academic...below. We’re happy to answer questions about this transition (so far) and what we’ve learned.  Thank you ...

  37. Truncated gRNAs for Regulating Gene Expression

    Type
    Blog Post
    Published
    Jan. 10, 2017, 3:37 p.m.
    ...nuclease-null—in order to effect different perturbations. Consider, for instance, the challenge of attempting to ...alterations can be exploited to quickly and inexpensively generate a potent Cas9-based transcriptional...

  38. Plasmids 101: Knockout/Knock-In Plasmids

    Type
    Blog Post
    Published
    Dec. 1, 2016, 3:30 p.m.
    ...can survive positive selection, but it will be sensitive to negative selection. Remove the positive selection...creating knockout animals or cell lines, please consider sharing them by depositing with Addgene! Figures...

  39. 10 Basic tips for mammalian cell culture

    Type
    Blog Post
    Published
    July 12, 2018, 1:09 p.m.
    ...normally lies around 7.35-7.4 so mammalian cells are sensitive to pH. Anything between orange and light orange...Additional passages will allow your cells to achieve a consistent growth rate. For every minute spent organizing...

  40. Mobile-CRISPRi: Bringing CRISPRi to Diverse Bacteria

    Type
    Blog Post
    Published
    April 4, 2019, 12:53 p.m.
    ..., cultures treated with a folA gRNA were more sensitive to trimethoprim when compared to control gRNA ...for trimethoprim, there would be no changes in sensitivity. Pooled knockdowns are also possible with Mobile-CRISPRi...

  41. Tips for a 1st time AAV user (by a Rookie AAV user)

    Type
    Blog Post
    Published
    Oct. 23, 2018, 11:49 a.m.
    ...primarily found in the culture media, you should consider performing PEG precipitation. Instead of sonicating...solutions are carefully layered and then the viral suspension generated during AAV production is overlaid. ...

  42. Degrading DNA with Cascade-Cas3

    Type
    Blog Post
    Published
    Feb. 11, 2025, 2:15 p.m.
    ... increase efficiency of these systems, though consistent expression can still be a challenge. A multitasking...powerful than it is. The nuclease domain of Cas3 is considered weak, especially compared to the power of Cas9...

  43. Antibodies 101: Flow Compensation

    Type
    Blog Post
    Published
    May 16, 2024, 1:15 p.m.
    ...which way your population needs to be shifted, consider your perpendicular lines. In this case, the population...applying very large compensation values (100+), consider what two markers your fluorescent markers are ...

  44. Of Myc and Men

    Type
    Blog Post
    Published
    Jan. 19, 2023, 2:15 p.m.
    ...epitope, a phenomenon known as sequence context sensitivity (Schüchner, et al. 2020). In their study, some... newer Myc-tag antibodies appeared to be less sensitive to sequence context.    So, do Schüchner, et al...

  45. 3D Printing Meets CRISPR Cas9

    Type
    Blog Post
    Published
    Oct. 17, 2017, 12:55 p.m.
    ... 3-D path of the protein’s backbone in three-dimensional space, but their flexible and jiggly nature suggested...structure into experimentally determined electron density, have been known to say something silly like “See...

  46. Addgene's Cancer Collection Pages Connect the Research Community

    Type
    Blog Post
    Published
    Sept. 6, 2016, 2:30 p.m.
    ...suppressors, and their protein products. Proto-oncogenes consist of a group of genes/proteins that, when expressed...Ian A., Paul D. Lewis, and Carla Mattos. "A comprehensive survey of Ras mutations in cancer." Cancer research...

  47. Light Sheet Fluorescence Microscopy

    Type
    Blog Post
    Published
    July 16, 2015, 2:30 p.m.
    ... capture large field-of-views (6). For more comprehensive reviews on several other LSFM configurations...Bradke laboratories later developed the 3DISCO (3-dimensional imaging of solvent cleared organs) method of ...

  48. Tips for arabidopsis transformation

    Type
    Blog Post
    Published
    Oct. 25, 2018, 1:23 p.m.
    ...that is wide enough to dunk your plant’s flowers inside. 50 mL falcon tubes or plastic weigh boats are ...from a colony on a plate will reach an optimal density in 18-24 hours. I then add 0.5 mL of this starter...

  49. New Optimized Genome-wide CRISPRko, CRISPRi, and CRISPRa Libraries

    Type
    Blog Post
    Published
    Oct. 4, 2018, 12:44 p.m.
    ...human CRISPRko sgRNA library The Brunello library consists of 77,441 sgRNAs with an average of 4 sgRNAs per...Mediator) uses an improved activator design that consists of inactive Cas9-VP64, a sgRNA with two MS2 RNA...
Showing: 451 - 500 of 725 results