Skip to main content
Addgene
Showing: 221 - 240 of 706 results
  1. REPLACR Mutagenesis: Replacing In Vitro Recombination Methods

    Type
    Blog Post
    ...genomic SDM relatively straightforward, but plasmid-based SDM has lagged behind. While commercial kits are...PCR is used to splice together two PCR amplicons based on sequence homology, and the subsequent product...primers also have some homology to each other; ~17 bases is optimal. After being treated with DpnI to digest...
  2. Synthetic Photobiology: Optogenetics for E. coli

    Type
    Blog Post
    ...limited to neuroscience. In fact, precise light-based control of biological processes is highly useful...better fine-tune the level of output gene expression based on the input light intensity. In addition to these...strains of E. coli, as well as with other plasmid-based systems, which will enable the assembly of larger...
  3. Kiran Musunuru on the Newest TALEN Genome-Editing System

    Type
    Blog Post
    ...a DNA-binding domain to recognize any desired 15 base pair sequence in the genome in rapid fashion. The...All you do to create a protein that binds those 15 base pairs is take four plasmids out of the library in...genome-editing system for generating human stem cell-based disease models." Cell Stem Cell. 12, 238-251 (7 ...
  4. Alex Chavez on MD PhDs, CRISPR, Gene Drives, & More

    Type
    Blog Post
    ...and what work he's planning on doing with CRISPR-based gene drives focusing on safety and the ability to...Board with a particular interest in generating Cas9 based tools to enable facile control of DNA and RNA.   ...
  5. Transferable Skills Guide: Teamwork

    Type
    Blog Post
    ... knowledge base is going to necessarily need a diverse group of people to provide that base. Sometimes... Mission accomplished! One example of a mission-based team in academia would be your school’s post-doc...Let’s think about some of the advantages of team-based work: Nobody knows everything anymore John F. Kennedy... post, so here are just a few critical features based on my own experience: Leadership Few things are ...
  6. AAV Titers: Where Do They Come From And What Do They Mean?

    Type
    Blog Post
    ...infectious titer of a viral vector stock can vary based on transduction target and can also be altered by...titer is typically calculated by two popular PCR-based methods: Quantitative PCR (qPCR) Digital droplet...not widely used today (Fagone et al., 2012). PCR-based methods are robust, easy, fast, and convenient. ...quantified with a different primer pair. Ct values vary based on the amount of starting material in the PCR because...
  7. 3 Tips to Improve HDR Efficiency for CRISPR Editing in Human Cells

    Type
    Blog Post
    ... et al) we were very excited to set up a CRISPR-based editing platform in our lab. Our goal was to use...binding sequence, we recommend changing several bases as close to the PAM as possible (see below to find...Ludwig-Maximilians-University Munich. His lab develops iPSC-based models of neurodegenerative and neurovascular diseases...of Alzheimer’s disease by developing novel iPSC-based model systems.     References 1. Cong, Le, et al...
  8. Plasmids 101: Broad Host Range Plasmids

    Type
    Blog Post
    ...are functionally differentiated from one another. Based on their structural elements, oris interact with...proteins within different host organisms. pBBR1-based plasmids and plasmids in incompatibility groups ...campestris pv. malvacearum, Zymomonas mobilis pBBR1 based pBBR1  Gram-negative  Alcaligenes eutrophus...constitutive and IPTG-inducible integron promoter-based expression systems for Escherichia coli and Agrobacterium...
  9. Site Directed Mutagenesis by PCR

    Type
    Blog Post
    ...of the desired mutation (usually 1-3 mismatched bases) is sufficient for your primers to successfully ...a minor extension can usually ensure that the 3’-base(s) do not form secondary structures. The introduction...feature used in TA-cloning). Such non-complementary bases at the 3’-end interfere with plasmid reconstitution...et al. "Predicting DNA duplex stability from the base sequence." Proceedings of the National Academy of...
  10. Getting the Most from Your Lentiviral Transduction

    Type
    Blog Post
    ...the best possible results for any tissue culture-based experiment, labs should routinely test for mycoplasma...detector cell lines, ELISA, fluorometry, or PCR-based approaches to detect even low levels of contamination...detection, Uphoff et. al. outline a do it yourself PCR-based approach complete with freely available positive...concentration and titration of pseudotyped HIV-1-based lentiviral vectors."Nature protocols 4.4 (2009):...
  11. How to Negotiate a Successful Lab Start Up Budget

    Type
    Blog Post
    ... decisions based on experimental timelines and specific aims. 2. Then make decisions based on financial...instructions on creating a budget, making decisions based on that budget, and negotiating in regards to the...will need, and how long it will take to publish based on these findings. Most often we suggest estimating...
  12. Screening for Successful Genome Editing with Digital PCR

    Type
    Blog Post
    ...allows classification of individual target molecules based on their mutation status. The drop-off probe is ...References 1. Miyaoka, Y. et al. Isolation of single-base genome-edited human iPS cells without antibiotic..., Berman, J. R. & Postovit, L.-M. A Digital PCR-Based Method for Efficient and Highly Specific Screening...
  13. A Quick Guide to a Career in Software Product Management

    Type
    Blog Post
    ...contributed by guest blogger, Rachel Rubinstein, a field-based software product manager at Oxford Nanopore Technologies... messy if you’re not on top of it. In a science-based software PM role, having a good understanding of...Rubinstein! Rachel Rubinstein is currently a Field-based Product Manager at Oxford Nanopore Technologies ...
  14. Sharing is Caring: But How to Distribute Open Hardware?

    Type
    Blog Post
    ...of imaging data (ILASTIK), or the popular plugin-based image process framework FIJI are just three of the...to the biological protocol, and assembly designs based around the use of commercially available tools and...provides a potential conflict between traditional IP-based models and the use of taxpayer money to support ...
  15. Empty Backbones - Choosing Your Perfect Plasmid Backbone

    Type
    Collection
    ...Flag for bacterial expression pFA6a vectors - PCR-based C-terminal tagging in S. cerevisiae pCS2FLAG - N...expression pnCasPA-BEC - A cytidine deaminase-mediated base-editing plasmid in Pseudomonas species Vectors for...agroinfection-compatible Tobacco mosaic virus (TMV)-based transient expression vector GFP Varies pMKO.1 GFP...
Showing: 221 - 240 of 706 results