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Showing: 201 - 250 of 573 results

  1. A Look at Addgene's QC Process

    Type
    Blog Post
    Published
    May 6, 2025, 1:15 p.m.
    ...in SnapGene. SnapGene annotates the plasmid maps based on SnapGene’s “Common Feature” library — with some... the deposited scientist  The backbone elements based on the information provided and vector type  Any...assembly issues that arise due to the short 200–250 base pair reads sequenced on the Illumina MiSeq.  Resolving...

  2. Plasmids 101: The Wide World of Natural Plasmids

    Type
    Blog Post
    Published
    Oct. 8, 2024, 1:15 p.m.
    ...have proposed developing new cloning techniques based on these vectors (Shareck et al., 2004). Plasmid...al., 2023). In this post, we’ve grouped plasmids based on the phenotype they offer their host cell, but...Lee, B., & Miguez, C. B. (2004). Cloning Vectors Based on Cryptic Plasmids Isolated from Lactic Acid Bacteria...

  3. Progress Towards a PAM-Free CRISPR

    Type
    Blog Post
    Published
    March 7, 2024, 2:39 p.m.
    ...Cas9  NG  - SpRY nuclease  C to T Base Editor SpRY A to G Base Editor SpRY   NYN/NRN NYN is less ...SpRY was validated in double stranded cleavage and base editing applications, with high editing capacity...

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

    Type
    Blog Post
    Published
    Nov. 7, 2017, 1:59 p.m.
    ... that need them. Adeno-associated virus (AAV) – based delivery techniques may hold the key. In the simplest...effective. Challenges for AAV gene therapy Although AAV-based gene therapies are promising, scientists are working...should be taken into account when developing AAV-based therapies. Furthermore, deeper understanding of ...

  5. Hot Plasmids and Viral Preps - January 2021

    Type
    Blog Post
    Published
    Jan. 5, 2021, 2:15 p.m.
    ...Francois St-Pierre’s lab developed a new form of FACS-based cell screening, Single-cell Phenotypic Observation...Plasmids and Resources page.  CDDR (CRISPR–Cas9-based Dual-fluorescent DSB Repair) is a double-strand ...double-strand break repair assay for mammalian cells. It's based on the introduction & resolution of two DSBs in a...eliminate competing mobile genetic elements. Image by Basem Al-Shayeb in the Doudna Lab. Scientists looking...

  6. Plasmids 101: 5 factors to help you choose the right cloning method

    Type
    Blog Post
    Published
    Aug. 21, 2018, 12:31 p.m.
    ... you. Traditional restriction enzyme and ligase based cloning is a relatively inexpensive way to get the...a try. This method, developed by Invitrogen and based on lambda phage integration sequences, relies on...inserts a breeze. Among them are some of the homology based methods, like Gibson or SLIC, since one can pool...

  7. Plasmids 101: Optimizing Plasmid Yields

    Type
    Blog Post
    Published
    May 26, 2016, 2:30 p.m.
    ...viewed more as a guideline, since they are typically based on the copy number of the empty plasmid backbone...matrix could clog the matrix when performing filter based plasmid prepping. Be sure to stay within the recommended...for each plasmid/strain combination individually, based on determining the cell density or by harvesting...

  8. Plasmids 101: Terminators and PolyA signals

    Type
    Blog Post
    Published
    March 31, 2016, 2:30 p.m.
    ...two broad categories, prokaryotic or eukaryotic, based on the functional elements they contain. Plasmid...polyadenylation? The role of the terminator, a sequence-based element, is to define the end of a transcriptional...terminators are not usually employed in plasmid-based expression systems, so these will not be detailed...

  9. Our Most Popular Blog Posts from 2019

    Type
    Blog Post
    Published
    Dec. 31, 2019, 2:26 p.m.
    ...introduce insertions, deletions, and all possible base-to-base changes.   1. Plasmids 101: Transformation,...

  10. Mycoplasma Contamination: Where Does It Come From and How to Prevent It

    Type
    Blog Post
    Published
    Oct. 8, 2020, 1:15 p.m.
    ...know it’s there? For this task, you can use PCR based detection, DNA staining, fluorescent tagging, agar...and cost-effective (Sung, 2006). The sensor cell-based method uses specialized cells that can detect the...from different biotech companies which are enzyme-based mycoplasma detection kits. These kits detect mycoplasma...YJ, Hong JT, Chung YB, Lee CK, Song S (2006) PCR-based detection of mycoplasma species. J Microbiol .44...

  11. PCR Applications for Copy Number Variation Assays

    Type
    Blog Post
    Published
    April 26, 2022, 1:15 p.m.
    ... in this blog post, we're going to focus on PCR-based applications (qPCR and ddPCR.) qPCR Quantitative.... Quantitative analysis of copy number variants based on real-time LightCycler PCR. Curr Protoc Hum Genet...genetic elements can be determined by SYBR-Green-based quantitative real-time PCR. J Microbiol Methods....sample with known copy number set as the expression baseline. Copy number is then determined by comparing results...

  12. Fluorescent Tagging of Endogenous Genes with SapTrap

    Type
    Blog Post
    Published
    Oct. 12, 2017, 2:26 p.m.
    ...lab developed a polymerase chain reaction (PCR)-based system that allows users to generate a plasmid containing...Masato Kanemaki's lab has developed a CRISPR/Cas-based system for tagging endogenous proteins with an auxin-inducible...Schmid-Burgk, Jonathan L., et al. “CRISPaint allows modular base-specific gene tagging using a ligase-4-dependent...

  13. Google Forums Round Up: First Impressions of NgAgo

    Type
    Blog Post
    Published
    Aug. 4, 2016, 2:30 p.m.
    ...experiences with the new method and look for advice/tips. Based on the discussions in the forums, it appeared that...majority of the respondents (70%) used a lipofectamine-based method to deliver NgAgo ingredients into the cell.... Overall, the real take-home message thus far, based on survey respondents "Final Verdict", is that more...

  14. The Open Repository of CRISPR Screens: CRISPR Screen Data in One Place

    Type
    Blog Post
    Published
    Jan. 21, 2020, 2:30 p.m.
    ...in order to increase the accessibility of CRISPR-based phenotypic screen datasets. Original published CRISPR...demonstrated the feasibility and power of CRISPR-based library screens in cell lines from humans and other...? The Open Repository of CRISPR Screens (ORCS) database is a new freely available resource developed by...which can also be used to search the entire CRISPR database for particular screens of interest.   Figure...screen data included in the BioGRID ORCS CRISPR database are encouraged to contact BioGRID curators at ...

  15. X-CHIME: Context Dependent Germline Knockout in Immune Cells

    Type
    Blog Post
    Published
    Jan. 16, 2024, 6:13 p.m.
    ..., as it only changes one gene at a time. CRISPR-based systems enable gene knockout in immune cells in ...out a single gene in defined cell types (lineage-based) pXPR_070 Violet-excited GFP (vex) Lineage-Cre...

  18. What’s in Your Tube?: A Quick Guide to Measuring DNA by Spectrophotometry

    Type
    Blog Post
    Published
    July 1, 2025, 1:15 p.m.
    ...the most popular options, so most of this post is based on reading results from a NanoDrop. The principles...calculate a sample’s nucleic acid concentration based on the sample’s absorption at 260 nm (A260). ...use a complementary assay, such as a fluorescence based assay or running an agarose gel. In the case of ...

  19. Degrading DNA with Cascade-Cas3

    Type
    Blog Post
    Published
    Feb. 11, 2025, 2:15 p.m.
    ...number of ways, from small edits that change single base pairs, to chromosomal inversions and large deletions...1 CRISPR systems make up the majority of CRISPR-based immune systems in bacteria and archaea. Class 1 ...deletions of up to hundreds or even thousands of base pairs, stopping either at boundaries defined by ...originally suspected that Cas3 moved a couple of kilobases down the strand of DNA, Cascade caught up to it...

  20. Career Insights: Technical Support Specialist

    Type
    Blog Post
    Published
    Nov. 9, 2017, 2 p.m.
    ..., technical support is often covered by the lab based team as part of their general job duties. Bigger...etc. Technical support roles are primarily office based, and email/phone are the most popular ways to communicate... pressure which are often found in sales or lab-based roles in industry. What to look forward to in a ...answers for customers, internal resources and databases are scrutinized, and this information is enriched...

  21. New CRISPR Tools: Cas7-11 and PASTE

    Type
    Blog Post
    Published
    Feb. 17, 2022, 2 p.m.
    ...would represent a major step forward for CRISPR-based RNA editing - and further experiments strongly supported...CRISPR could cut DNA, the second generation could do base editing, and with the third generation came Prime...ability to now make any small changes, including base edits, insertions, and deletions. But CRISPR’s uses...deletions and small insertions, leaving a several kilobase-sized gap in the gene editing field. The AbuGoot...now developed PASTE, which can “drag and drop” kilobases’ worth of DNA with CRISPR-driven precision into...

  22. CRISPR Between the Genes: How to Experiment with Enhancers and Epigenomics

    Type
    Blog Post
    Published
    Aug. 9, 2016, 2:30 p.m.
    ... the very beginning of the third chromosome, ten base pairs in length. An emerging family of sequencing...increase the expression of a gene that is millions of base pairs away. (For details on DNA compaction and the...parsed for spCas9 PAM sites (NGG) and then filtered based on two main criteria: no TTTTT allowed (this is ...here with permission. Quilt is a collaborative database for genomics. In this article, Quilt CTO Aneesh...is CTO at Quilt Data. Quilt is a collaborative database. Aneesh's research interests span machine learning...

  23. Tips for Screening with Yeast Two Hybrid Systems

    Type
    Blog Post
    Published
    Oct. 22, 2015, 2:30 p.m.
    ...common to use many preys with a single bait in Y2H-based screens. In this way, proteins that interact with...proteome of a particular cell type, or a subset based on a cellular compartment or growth stage). A prey...protein-protein interactions in mammalian cells based on protein splicing. Anal Chem. 2001; 73:2516-2521... Including a prey-only control also provides a baseline level of reporter activation. Vary the expression...

  24. New Antibodies in the Lab Animation!

    Type
    Blog Post
    Published
    Nov. 19, 2024, 2:15 p.m.
    ...Thinking about using antibody-based applications in your experiment...but not sure where to start or ...

  25. Antibodies 101: Introduction to Antibodies

    Type
    Blog Post
    Published
    Jan. 19, 2021, 2:15 p.m.
    ...immunoglobulin genes, recombinant antibodies are plasmid-based. Their sequences can be optimized for specificity...Arbabi-Ghahroudi et al., 2017). Although they’re based on the immune system of llamas, synthetic nanobodies...mixture of proteins. The proteins are separated based on molecular weight by SDS-PAGE, and are subsequently...well as tools for developing and producing plasmid-based antibodies - and we’re always growing our collection...

  26. Optogenetics + CRISPR, Using Light to Control Genome Editing

    Type
    Blog Post
    Published
    Sept. 3, 2020, 12:15 p.m.
    ...lab at Duke University developed similar systems based on the light-inducible heterodimerizing cryptochrome...systems have been developed for use with CRISPR-based gene editing (see table below). For example, the...possibilities for optogenetic manipulation of CRISPR-based systems in animal models. Use Anti-CRISPRs to Turn...Hasegawa S, Yoshimoto K, Sato M (2017) CRISPR–Cas9-based photoactivatable transcription systems to induce..., Kawano F, Suzuki H, Sato M (2015) CRISPR-Cas9-based Photoactivatable Transcription System. Chemistry...transcription Zhou et al., 2020   NPOM-caged nucleobases Cas9, photocaged-gRNA Gene editing For more...

  27. 28 Hot Plasmid Technologies from 2015

    Type
    Blog Post
    Published
    Dec. 23, 2015, 3:30 p.m.
    ... Santalucía Lab has developed a cloning toolkit based on the Multisite Gateway® technology from Life Technologies...MultiSite Gateway Kit is a customizable Gateway®-based toolkit that allows scientists to generate fusion... Medical School has recently developed a CRISPR-based multicolor tool enabling the imaging of multiple...cancer-associated chromosome shattering. This method is based on the recognition of genomic loci by three orthogonal...does it work? RUSH is an ingenious two-state assay based on the reversible interaction between a hook protein...Neveu laboratory designed a new iterative, chaining-based cloning method that simplifies the process of constructing...presented Archaerhodopsin 3 (Arch) as a new rhodopsin based voltage indicator for use in the detection of neuronal...

  28. Genetically-encoded Sparse Cell Labeling - A SPARC of Innovation

    Type
    Blog Post
    Published
    May 21, 2020, 1:15 p.m.
    ...tools to overcome this restraint include an AAV-based sparse labeling system where a limiting amount of... truncated attP site. The three SPARC variants, based on the length of the attP site, allow for dense ...

  29. Addgene Begins Distribution of Recombinant Antibodies

    Type
    Blog Post
    Published
    March 28, 2022, 4:30 p.m.
    ... 2015). All of Addgene’s antibodies are plasmid-based and recombinant. Learn more about recombinant antibodies...affinity reagents, such as nanobodies and ScFvs, based on community demand. Figure 1: Antibody...from Addgene via our Antibody Data Hub once the database is available. This data will help the collection...

  31. Using Addgene's New Viral Service for Your Research

    Type
    Blog Post
    Published
    Nov. 17, 2016, 3:30 p.m.
    ... time, we’re developing ourselves as a knowledge base for the processes of viral production and testing...connections. Expanding the viral vector knowledge base: You can help! We’ve been expanding our viral vector...

  32. Extracurricular Activities for a Strong Science Career Path

    Type
    Blog Post
    Published
    Jan. 21, 2014, 3:08 p.m.
    ...alternative careers. Science Careers has developed a web-based, career-planning platform called the Individual ...to learn about a variety of science professions based on their skills and interests. Once scientists identify...

  33. Antibodies 101: Introduction to Immunofluorescence

    Type
    Blog Post
    Published
    Nov. 16, 2021, 2:15 p.m.
    ...found in my tissue.” It is based on the same principles as other antibody-based assays, like an ELISA or...cases, a whole organism. But, as with any antibody-based technique, validation and proper experimental design...different companies, such as Validated Antibody Database and CiteAb. The antibody you choose can make or...

  34. Viral Vectors 101: Transductions

    Type
    Blog Post
    Published
    April 30, 2024, 1:15 p.m.
    ...long-term (AAV). If you perform a lipid-based or electrical-based transfection, most of the nucleic acid...

  35. In Vivo Biotinylation of Bacterial Fusion Proteins

    Type
    Blog Post
    Published
    Jan. 25, 2018, 2:09 p.m.
    ...interaction approaches that of GST-glutathione. Biotin-based immobilization for kinetic analysis Proteolytic ...bacteria. Our initial attempt to create a plasmid based on the biotin ligase BirA backbone (Eric Campeau...

  36. 25 Work From Home Tips for Scientists

    Type
    Blog Post
    Published
    March 19, 2020, 1:15 p.m.
    ... remotely can be more difficult if your work is based in the lab. If you’re unsure what to do during this...submitting a registered report, where peer review is based on the methods, not the results. Also consider using...

  37. Tetbow: Bright Multicolor Labeling for Neuronal Tracing

    Type
    Blog Post
    Published
    Jan. 24, 2019, 2:24 p.m.
    ...discriminating between neurons for light microscopy-based neuronal reconstruction. To achieve stochastic multicolor...conventional transgenic strategies. In such vector-based gene delivery systems, the stochastic expression...

  38. "Build Your Plasmid": The Game - Play to Teach

    Type
    Blog Post
    Published
    July 17, 2018, 12:23 p.m.
    ...should be in real scientific practice, so it is based on a win-win strategy. After each team presented...developed to teach gamma-retrovirus or lentivirus-based ex-vivo gene therapy’ strategies to undergraduates...

  40. Plasmids 101: TOPO Cloning

    Type
    Blog Post
    Published
    Oct. 27, 2016, 2:30 p.m.
    ...Topoisomerase based cloning (TOPO cloning) is a DNA cloning method that does not use restriction enzymes...polymerase has an error rate of about 1 in 3,500 bases. Typically polymerases with proofreading functionality... relies on the basic ability of complementary basepairs adenine (A) and thymine (T) to hybridize and form...

  42. Antibodies 101: Beyond Surface Labeling

    Type
    Blog Post
    Published
    June 27, 2024, 1:15 p.m.
    ... MACS. But MACS usually only allows for sorting based on a single (lineage) marker, while fluorescence-activated...biorender.com.   Dye stains Dye stains are non-antibody-based stains that bind DNA or free amines of proteins....cytosolic and/or nuclear markers. Lastly, stains, based not on antibodies but on small molecule dyes, are...

  43. An Integrin Antibody Toolkit from IPI

    Type
    Blog Post
    Published
    March 28, 2024, 1:15 p.m.
    ...Institute for Protein Innovation (IPI), a Boston-based, nonprofit protein research institute, partnered...unconjugated and recommended for use in flow cytometry. Based on the established applications of the original ...with the Boston Children’s Hospital and Harvard-based lab of Tim Springer, the founder of IPI. Caitlin...

  44. Hot Biosensors 2022: Year-End Roundup

    Type
    Blog Post
    Published
    Dec. 6, 2022, 2:15 p.m.
    ...They also produce more than 20 other MTRIA sensors based on GPCRs recognizing various neurotransmitters, ...powerful high-throughput screen to select variants based on performance metrics specifically for two-photon...phosphorylated state of an immunoreceptor tyrosinase-based activation motif (ITAM). By fusing ITAM to an RTK...

  45. Antibodies 101: Single Chain Fragment Variables (scFvs)

    Type
    Blog Post
    Published
    June 3, 2021, 1:15 p.m.
    ... The HA frankenbody works just like an antibody-based probe, but is easier and cheaper to generate since...scFvs: Just like antibodies, there are many scFv-based therapies in clinical trials or already in the clinic...blog: Read about frankenbodies, an anti-HA scFv-based probe for fluorescent imaging. Learn more about ...

  46. CRISPR 101: Off-Target Effects

    Type
    Blog Post
    Published
    Feb. 8, 2024, 2:15 p.m.
    ...with the bulkiness that accompanies mispairing of bases. To reduce off-targets, you’ll want to select a ... Off-targets that come from mutations arise from base/prime editing or Cas cleavage. In the case of Cas...Resources   References Rabinowitz R, Offen D. Single-Base Resolution: Increasing the Specificity of the CRISPR-Cas...

  47. New Optogenetic Tools for Cytoskeleton and Membrane Control

    Type
    Blog Post
    Published
    Sept. 28, 2023, 1:15 p.m.
    ... optogenetic tools for manipulating Rho GTPases based on BcLOV4. These single-component systems take advantage...interested in directed-evolution methods, the activity-based selection in mammalian cells used here could be ...Fromme, J. C., & Baskin, J. M. (2023). Activity-based directed evolution of a membrane editor in mammalian...

  48. Evolution of Brainbow: Using Cre-lox for Multicolor Labeling of Neurons

    Type
    Blog Post
    Published
    April 24, 2015, 2:39 p.m.
    ..., especially in in vivo models, and new Cre-lox based technologies are still being developed today. In...Flpbow construct that is functionally similar to Cre-based Brainbow, but is controlled by FLP/FRT recombinase...-44. PubMed. Drosophila Brainbow: a recombinase-based fluorescence labeling technique to subdivide neural...

  49. A Novice's Perspective on Neuroscience - SfN2017

    Type
    Blog Post
    Published
    Dec. 15, 2017, 2:07 p.m.
    ...to altered function or maybe your favorite FRET-based biosensing technique can be applied to an important...experiments enabled by physical, software, and biology-based tools as well as collaborations between different...
Showing: 201 - 250 of 573 results