Skip to main content

We narrowed to 373 results for: proc.2

Showing: 251 - 300 of 373 results
  1. How to Deposit Your Plasmids with Addgene

    Type
    Blog Post
    Published
    Feb. 27, 2024, 2:15 p.m.
    ...two options to begin your plasmid deposit. Figure 2: Choose to submit plasmid(s) online or by using a ...format. Figure 6: Add sequence, maps, and files. Step 2: Gene and Insert *Note: This page will not appear ...starting the process. After you have logged in to your account, you can begin the deposit process by clicking... as a step-by-step guide to the online deposit process. To deposit plasmids online, you will need to create...the Deposit menu: Figure 1: Begin the deposit process by clicking "Submit Online Deposit" or "Download...published in a peer-reviewed journal. To start this process, simply enter your preprint title or a general ...Hold for publication: we will initiate our MTA process and perform standard quality control but will not...
  2. Viral Genome Sequencing - A One Stop Shop for AAV Identity and Serotype Confirmation

    Type
    Blog Post
    Published
    April 23, 2020, 1:15 p.m.
    ...containing complementary polynucleotide chains. J Virol 9(2): 394-396. Lecomte E, Tournaire B, Cogné B, Dupont...   While the VGS process isn’t new to Addgene, we are excited to share the procedures and Python scripts... vector (AAV) samples and created a simplified process, termed viral genome sequencing (VGS), to confirm...We recently published a description of the VGS process and use cases in Human Gene Therapy. Existing methods...substrates for tagmentation-based library preparation procedures. This finding eliminates the double-stranding...
  3. Plasmids 101: Gram Negative and Positive Bacteria

    Type
    Blog Post
    Published
    Aug. 2, 2022, 1:15 p.m.
    ...Bacterial Cell Envelope. Cold Spring Harb Perspect Biol. 2(5) a000414 (2010). Cleveland Clinic. Gram Stain. https...bacteria’s thinner membrane. Procedure The stain is a pretty simple process. First, flood the heat-fixed...pneumoniae, while the alcohol used in the staining process bleached another, Klebsiella pneumoniae. After ...photos/occbio/7092385559 For more specific procedural instructions, including timing, preparing the... them red or pink. At the end of the staining procedure, gram positive bacteria are stained purple or ...
  4. The Power Behind NGS Plasmid Validation: seqWell

    Type
    Blog Post
    Published
    April 19, 2017, 3:25 p.m.
    ...from a single Illumina MiSeq® run. As shown in Fig. 2, below, differences in input concentration across ...laboratories from using these methods for routine processes like plasmid validation. At seqWell, Inc., our...sample-specific barcodes. Rather than continuing to process 100s to 1000s of samples individually, plexWell...additional benefit of plexWell™’s sequential tagging procedure, the quantities of library molecules for each ...a wide range of input DNA concentrations. This process is explained more fully in the animated Fig. 1 ...samples are brought in line with each other before proceeding to library prep. Even with this “pre-prep” normalization...
  5. Sequencing Options for CRISPR Genotyping

    Type
    Blog Post
    Published
    Oct. 4, 2016, 2:30 p.m.
    ...closer to the clinic (Tsai and Joung 2016). Table 2: Unbiased Genotyping Options Technique Detection...PMID: 23792628. PubMed Central PMCID: PMC3773023. 2. Vouillot, Léna, Aurore Thélie, and Nicolas Pollet...most important steps in the CRISPR experimental process is validating edits. Regardless of which CRISPR...not an edit in the target gene. The validation process, also known as CRISPR genotyping, is critical to...
  6. Antibodies 101: Fc Effector Functions

    Type
    Blog Post
    Published
    May 18, 2023, 1:15 p.m.
    ...affinity and specificity. Trends in Immunology, 29(2), 91–97. https://doi.org/10.1016/j.it.2007.11.004 ...mark it as a target for other immune cells, in a process known as opsonization. In antibody-dependent complement... the pathogen the antibodies are bound to in a process known as phagocytosis.  Other functions We’re still...
  7. Bricking Science: Portraying Scientific Reality Through LEGO

    Type
    Blog Post
    Published
    Nov. 29, 2016, 3:30 p.m.
    ...motion videos is very time consuming; producing a 1-2 minute video takes an entire week of our free time...one step. Challenges in the LEGO science video process One major challenge when we started was creating...laboratory. When we started, the hardest part of the process was to embed personality in pieces of plastic, ...
  8. Six Spooky Science Stories and Halloween at Addgene

    Type
    Blog Post
    Published
    Oct. 31, 2019, 1:06 p.m.
    ...melanogaster." Molecular and cellular endocrinology 215.1-2 (2004): 1-10. PubMed PMID: 15026169. Soo, Rochelle...harbouring hairworm: a proteomics approach." Proceedings of the Royal Society B: Biological Sciences 272.1577...
  9. Viral Vectors 101: Transductions

    Type
    Blog Post
    Published
    April 30, 2024, 1:15 p.m.
    ...strategy, etc.) and episomal expression will last ~2-7 days at moderate levels. With regard to target cells... What is a transduction?  Transduction is the process of transferring foreign nucleic acid into a cell.... Transductions are similar to transfections, processes which introduce foreign nucleic acid into cells...   Figure 1: Transduction to expression process. A viral particle enters a host cell, enters the...
  10. Don’t Wait To Launch Your Health Science Startup

    Type
    Blog Post
    Published
    Aug. 25, 2015, 2:30 p.m.
    ...yourself greater chances for success.  Build a team of 2 or 3 Don’t go it alone.  You’ll find that no matter...background and/or connections to even begin this process. I am the cofounder and CEO of miLab, a Boston-...innovative ideas in all facets of the entrepreneurial process in order to develop those ideas into commercial...people for guidance.  Before even beginning the process of launching a company, you need to determine whether...
  11. How Addgene Creates Content

    Type
    Blog Post
    Published
    Nov. 17, 2022, 2:15 p.m.
    ...and in line with Addgene style.        Fig. 2: Reference image (left) provided to our graphic designer...content. Regardless of the process, everything goes through an editing process and final check before publication...invite you behind the scenes in our content writing process, and to even get involved yourself. You probably...science, and our entire mission is to enable that process. Our repository of plasmids, AAVs, antibodies, ...recommendations. A few discussions in the review process led to quick dives back into the literature to ...and hit publish. Making content is an iterative process. We are constantly reviewing the resources we currently...
  12. Antibodies 101: Validation

    Type
    Blog Post
    Published
    March 24, 2022, 1:15 p.m.
    ...another. You’ll need to validate that points 1 and 2 hold true under your specific experimental conditions...when (1) other strategies may not be feasible and (2) large-scale proteomic and transcriptomic datasets...identified as your target protein, allowing you to proceed with confidence.  Share your results Ultimately...
  13. Finding Your Science Policy Path

    Type
    Blog Post
    Published
    Sept. 22, 2020, 1:15 p.m.
    ...and during your application process so you can learn more! Figure 2: The many ways to prepare for...teaching and mentoring others (see more in Figure 2). Starting early demonstrates not only your interest...
  14. What's the Best Way to Elute and Store Your Plasmid DNA?

    Type
    Blog Post
    Published
    June 4, 2020, 1:22 p.m.
    ...elution choices: TE, Tris buffer, or water Figure 2: Tris-EDTA, Tris, or water? The buffer you choose ...the plasmid, and still allowing most downstream processing. On the other hand, nuclease free water is a ...
  15. Components of CRISPR/Cas9

    Type
    Blog Post
    Published
    Feb. 2, 2016, 5 p.m.
    ...Without binding to the guide RNA, Cas9 cannot cut. 2. Bind to target DNA in the presence of a guide RNA...systems have a variety of enzymes responsible for processing foreign DNA as well as the RNA guides required...scaffold linking the crRNA to Cas9 and facilitates processing of mature crRNAs from pre-crRNAs derived from...repeat regions contain sequences required for processing pre-crRNA into mature crRNA and tracrRNA binding... Overview of native CRISPR arrays and their processing for cleaving foreign DNA. A) CRISPR arrays as...
  16. Cancer and the Immune System: Deciphering the Relationship

    Type
    Blog Post
    Published
    March 14, 2017, 2:30 p.m.
    ...IFNγ) - and eliminate cancer cells. 2. Equilibrium - In a process analogous to the development of drug... 35. Academic Press, 2015. PubMed PMID: 25818339. 2. Curiel, Tyler J., et al. "Specific recruitment of...system, tumors “learn” how to escape it. In this process, cancer cells develop and thrive within what is...
  17. Three Tips for Preventing Viral Plasmid Recombination in Your Samples

    Type
    Blog Post
    Published
    Sept. 14, 2021, 1:15 p.m.
    ...extraction protocol for more details.   Figure 2: An uncut plasmid is expected to form 3 bands: nicked...intramolecular DNA recombination. This naturally occurring process is mediated by recombinases like RecA and RecBCD...analysis of in vitro intramolecular recombination. Proceedings of the National Academy of Sciences 77:4847–4851...
  18. 5 Uncommon Ways to Gain Career Skills During an Uncommon Time

    Type
    Blog Post
    Published
    Oct. 20, 2020, 1:15 p.m.
    ...peer-mentoring groups at Addgene have gone virtual!   2. Practice public speaking For almost any science career...their resumes have a definite advantage in our process. We see candidates who have done the following ...still be practicing and getting familiar with the process. Bonus points for being able to just look at a ...
  19. Creating accessible biology activities in schools with BioBits

    Type
    Blog Post
    Published
    Aug. 2, 2018, 12:56 p.m.
    ...PMID: 30083608. PubMed Central PMCID: PMC6070312. 2. Stark, Jessica C., et al. "BioBits™ Bright: A fluorescent...to be incorporated into a host cell (through a process called transformation), essentially giving the ...shelf-stable until water is added and protein production proceeds. FD-CF pellets are easy for students to use in...
  20. Sharing is Caring: But How to Distribute Open Hardware?

    Type
    Blog Post
    Published
    Feb. 28, 2023, 2:15 p.m.
    ... high-priced commercial devices.     Fig. 2: Workflow for open software (left) vs. open hardware...constantly being developed which combine computational processing with biological protocols and revolutionary physical...Software In the field of microscopy, where image processing algorithms for analyzing time-lapse images, quantitatively...data (ILASTIK), or the popular plugin-based image process framework FIJI are just three of the constantly...constant updates of the software. In hardware, this process is often much more involved. Challenges such as...these limitations into account in the development process. Widely used, commercially available tools such... the world that are otherwise cut off from the process.   About the author: Benedict Diederich received...
  21. Antibody Validation for Flow Cytometry

    Type
    Blog Post
    Published
    Sept. 10, 2024, 1:15 p.m.
    ...types within a sample to expected labeling (Figure 2). The panel of antibodies used in this approach should...the antibody in the same tube.     Figure 2: Schematic showing the use of an orthogonal approach...sample and target type. To start the validation process, one must first confirm that a candidate antibody...approved antibody clones The antibody validation process represents a high level of effort and time, which.... We find it useful to start with a screening procedure in easily genetically modified cell types (top... C. (2023). Scaling of an antibody validation procedure enables quantification of antibody performance...Implementation of an antibody characterization procedure and application to the major ALS/FTD disease gene...
  22. Addgene's Tips for Plasmid Quality Control

    Type
    Blog Post
    Published
    Jan. 14, 2016, 3:30 p.m.
    ... and transfer 100 µl of glycerol stock to each of 2 96-well microplates (identical copies, see image above...plasmids we distribute through our quality control processes. Every plasmid we receive is rigorously verified...every plasmid we distribute through our deposit process, and then we carefully track samples through our... to find all of the information needed for the process. Always sequence verify Once the physical sample...sequencing If you’re like us, you need a strategy for processing several sequencing reactions in parallel. We ...We have a few tricks to simplify the process. First, nearly all of our Sanger sequencing is done from glycerol...
  23. The CRISPR Software Matchmaker: A New Tool for Choosing the Best CRISPR Software for Your Needs

    Type
    Blog Post
    Published
    Nov. 3, 2015, 3:30 p.m.
    ... tabular HTML formatted output. Surprisingly only 2 tools provide FASTA output. A few notable tools allow...Nature biotechnology(2014). PubMed PMID: 24880488. 2. Hsu, Patrick D., et al. "DNA targeting specificity...development has caught up with it. The peer review process has been challenged with many closely spaced CRISPR... are trying to solve, and finally how we might proceed in the future. When we think of designing experiments...These functions help speed up the sgRNA design process by removing repetitive tasks and/or by providing...providing features to help with the post-design process such as primer and plasmid design. Examples:  “multiplex...
  24. CRISPR 101: Drug discovery and CRISPR

    Type
    Blog Post
    Published
    May 4, 2023, 1 p.m.
    ...development and therapy. Nat Rev Drug Discov (2017) 16(2):89-100. DOI: 10.1038/nrd.2016.238 Additional resources... into this workflow? CRISPR both speeds up the process and can help weed out a lot of poor drugs before...
  25. Sensing Neuronal Dopamine

    Type
    Blog Post
    Published
    July 30, 2019, 12:59 p.m.
    ...developed fluorescent-protein based DA sensors (Figure 2). The dLight1 dopamine sensor In 2018, Lin Tian’s ... its similarity to the “dopamine receptor”." Proceedings of the National Academy of Sciences 69.8 (1972...
  26. Understanding GPCR Signaling Using TRUPATH

    Type
    Blog Post
    Published
    Dec. 14, 2020, 2:15 p.m.
    ...dissociation by bioluminescence resonance energy transfer 2, or BRET2. With binding of the drug, the alpha and..., receptors that play many roles in signaling processes including smell, taste, sight, inflammation, and...
  27. All in a Twist: dsRNA

    Type
    Blog Post
    Published
    Feb. 2, 2023, 2:15 p.m.
    ... would release them into the cytosol. Fig. 2: Endogenous sources of duplexed RNA DNA:RNA hybrids... as part of their infection and/or replication process within the host. dsRNA viruses are inherently double...intermediates as part of the viral replication process. dsDNA viruses yield dsRNA through a slightly different...inherently generating RNA:DNA intermediates in the process. These transcription-coupled R-loops are transient...above play distinct roles in promoting cellular processes and functions, but they also have a darker side...
  28. Safe Port in a Storm...How Addgene is Weathering the Pandemic

    Type
    Blog Post
    Published
    Sept. 17, 2020, 1:15 p.m.
    ...majority of deposited materials to study SARS-CoV-2 would be fully open and available to industry requestors...product. Our daily operations for all ongoing processes are entirely self-supporting. We hire more staff...
  29. Why Add Sucrose? Improved Yields for Adeno-associated Virus Preparation

    Type
    Blog Post
    Published
    Dec. 13, 2018, 1:14 p.m.
    ...bioengineering 94.2 (2006): 322-329. PubMed PMID: 16528756. 2. Oh, Steve KW, et al. "Substantial overproduction ...Advances in animal cell biology and technology for bioprocesses (1989): 272-278. 4. Ozturk, Sadettin S., and...
  30. Addgene's Top 10 Blog Posts of 2015

    Type
    Blog Post
    Published
    Dec. 7, 2015, 6 p.m.
    ...HDR) and how it’s used in CRISPR genome editing.   2. Plasmids 101: Cre-lox Cre recombinase can be used...guides us through the basics of the validation process for CRISPR mediated genome engineering. 10. Plasmids...
  31. Extracurricular Activities for a Strong Science Career Path

    Type
    Blog Post
    Published
    Jan. 21, 2014, 3:08 p.m.
    ... sets Taking the time to identify and research 2 – 3 potential professions will highlight which skills...office positions   Market analyses Patent process Business Models Communication Regulatory ...
  32. Custom CRISPR Screens & the Green Listed Software

    Type
    Blog Post
    Published
    July 11, 2017, 2:30 p.m.
    ...six different research constellations: Doench/Root (2), Zhang (3), Wu (4), Yusa (5), Wang/Lander/Sabatini...Bioinformatics 33.7 (2016): 1099-1100. PubMed PMID: 28414855. 2. Doench, John G., et al. "Optimized sgRNA design to...genes, Green Listed can help make the gRNA design process quick and easy. Green Listed - a CRISPR Screen ...
  33. Using CRISPR/Cas9 to Edit Disease Out of the Genome

    Type
    Blog Post
    Published
    June 23, 2022, 1:15 p.m.
    ...cellular physiology.  https://doi.org/10.1002/jcp.30064 2. Barman et al. 2020. “A glance at genome editing with... tested in clinical trials. It is speeding the process of the discovery/development of genetic targets...off-target effects is a major limitation. The procedure will also need to be perfected until the efficiency...
  34. Plasmids 101: Mammalian Vectors

    Type
    Blog Post
    Published
    March 25, 2014, 3:15 p.m.
    ... 293HEK Inhibits termination step of translation 2-10 ug/mL G418/Geneticin neo HeLa, NIH3T3, CHO, 293HEK...material (such as plasmids) into mammalian cells, a process called transfection. Transfection is somewhat comparable.... Since the DNA introduced in the transfection process is not integrated into the nuclear genome, in the...
  35. Plasmids 101: Methylation and Restriction Enzymes

    Type
    Blog Post
    Published
    June 30, 2016, 2:30 p.m.
    ...account when simulating a digest as depicted in Lane 2. Controlling methylation Finally, you can control...protein expression. Methylases involved with these processes (for example Dam and Dcm methylases) are independent...used for site directed mutagenesis. During this process, incorporation of a desired mutation into your ...
  36. 3 Challenges in Plant Synthetic Biology

    Type
    Blog Post
    Published
    July 22, 2014, 5:46 p.m.
    ...and open a new conversation about GMOs.  Challenge #2: Technical obstacles to plant synthetic biology In...understand, and the complicated and unique metabolic processes of plants are being investigated on limited funding... developmental, metabolic, and photosynthetic processes. Often the tools developed for plant molecular...
  37. CRISPR/Cas9 FAQs Answered!

    Type
    Blog Post
    Published
    March 13, 2014, 4:08 p.m.
    ...extraction solution, runs the protocol, then uses around 2–4 µl in each 50 µl PCR reaction. The gDNA extracted... early onset Parkinson Point mutations. Cell, 146(2), 318–331. https://doi.org/10.1016/j.cell.2011.06.019...facilitate the processing of the crRNA) is also expressed from a separate promoter. In the pre-processing form,...construct from our 'Split RNA' design, where the pre-processing form of crRNA is expressed and a tracrRNA (the...in cell and with Cas9 and tracrRNA, it will be processed (i.e., cleaved) into a shorter form that contains...our 'Chimeric RNA' design (meaning the mature processed form of crRNA is fused chimerically to a mature...
  38. Plasmids 101: Sequence and Ligation Independent Cloning (SLIC)

    Type
    Blog Post
    Published
    Sept. 26, 2024, 1:15 p.m.
    ..., but at a lower efficiency (~20%). Figure 2: Schematic of multicomponent assembly with SLIC. Multiple...polymerize, allowing the slower exonuclease activity to proceed. By including only one type of dNTP in the reaction... can decide exactly how far the T4 exonuclease proceeds: at the first occurrence of the chosen nucleotide...: Schematic of SLIC. To start the SLIC cloning process, a gene of interest is PCR amplified to add 5′ ...
  39. Summer SciComm: Preprints

    Type
    Blog Post
    Published
    Aug. 16, 2022, 1:15 p.m.
    ... scientific communication ecosystem Figure 2: The scientific ecosystem, enriched by open access...These each have different goals, formats, and processes, and they provide different amounts of access ...journal-based peer review is usually a confidential process among a few individuals.  Early-career researchers...errors, with a lack of transparency in most review processes. Preprints and their feedback, in contrast, are...
  40. Visualizing Protein Turnover In Situ

    Type
    Blog Post
    Published
    Jan. 16, 2018, 3:20 p.m.
    ...PMID: 20543841. PubMed Central PMCID: PMC2920597. 2. Nathans, Daniel. "Puromycin inhibition of protein...effect of protein synthesis and degradation. These processes control the abundance of proteins in cells. Homeostatic...incorporation of puromycin into peptide chains." Proceedings of the National Academy of Sciences 51.4 (1964...puromycin of amino acid incorporation into protein." Proceedings of the National Academy of Sciences 45.12 (1959...
  41. Plasmids 101: Cre-lox

    Type
    Blog Post
    Published
    Jan. 13, 2015, 3:47 p.m.
    ..., N. and Hamilton, D. 1981. PubMed PMID: 6276558. 2. Site-specific DNA recombination in mammalian cells...the two strands are ligated, resulting in the reciprocal crossover event. This event can have three general...the stop codon is excised, and gene expression proceeds. One popular lox-stop-lox plasmid is from Tyler... remove the marker after the initial selection process. By floxing the selection marker, Cre can be used...
  42. How to Prepare for an Industry Interview

    Type
    Blog Post
    Published
    July 9, 2020, 1:15 p.m.
    ...company culture.  It’s also a good idea to prepare 1-2 unique questions for each interviewer by looking at...deliverables of your work, such as improving a process or procedure, solving a problem, or making a key decision... industry position is the on-site interviewing process. For both of my academic jobs, I interviewed with...different lab groups without having a formal interview process. Industry interviews, however, were more structured...lasted from half to all of the day.  This whole process may seem foreign and a little overwhelming, but...
  43. Multifunctional Neural Fibers: Modulation from Gut to Brain

    Type
    Blog Post
    Published
    Aug. 31, 2023, 1 p.m.
    ...delivering an AAV construct expressing channelrhodopsin 2 (ChR2), a blue light sensitive cation channel, fused...bells and whistles. The fibers are made from a process called thermal drawing, essentially where blocks...
  44. FPbase: A new community-editable fluorescent protein database

    Type
    Blog Post
    Published
    May 16, 2018, 1 p.m.
    ...incorporation of biosensors, protein evolution lineages, 2-photon spectra, more elaborate spectrum analysis tools...ways, and the advanced search page simplifies the process of building advanced multi-parametric search queries...
  45. Plasmids 101: NGS Quality Control for Pooled Libraries

    Type
    Blog Post
    Published
    Oct. 26, 2017, 1:59 p.m.
    ...created for the Brunello library is shown in Figure 2. You can use a small PCR product because the plasmids...talk about the amplification and verification processes we use to ensure high-quality library distribution...
  46. Resume Writing for Non-academic Science Careers

    Type
    Blog Post
    Published
    Dec. 18, 2014, 4:06 p.m.
    ... remember that a resume is typically no more than 2 pages long. Write your resume for the job posting ...familiar with the publishing or patent application processes? How about facilitating a collaboration between...
  47. Special Delivery: Fluorophore Targeting for FRET Studies

    Type
    Blog Post
    Published
    July 19, 2016, 2:30 p.m.
    ...establishing the dynamic range of your FRET assay (Figure 2). These labeling methods are freely interchangeable...PMID: 20161833. PubMed Central PMCID: PMC2802762. 2. Dumas, Anaëlle, et al. "Designing logical codon reassignment–Expanding...Ca2+ measurements using improved cameleons." Proceedings of the National Academy of Sciences 96.5 (1999...
Showing: 251 - 300 of 373 results