Skip to main content
Addgene

Enter a sequence to perform a BLAST-based search of the Addgene plasmid sequence database.

Information

In Category

We narrowed to 971 results for: LARS

Showing: 641 - 660 of 971 results

  1. Pouring LB Agar Plates

    Type
    Protocol
    ... you will be screening a large number of colonies, we recommend using larger plates. Many labs use 100... plates with colored marker - particular colors correspond to particular antibiotics. Position the flame...good idea of what that volume looks like in your particular plate. For the remainder of the plates, pour ...

  2. Plasmid Cloning by PCR (with Protocols)

    Type
    Protocol
    ...for PCR Based Cloning The basic PCR primers for molecular cloning consist of: Leader Sequence: Extra base...TAAGCAGAATTCATGTGGCATATCTCGAAGTAC-3'. For the Reverse Primer, the design is similar, but we need to use the reverse complement to get... get PCR amplification. We can start similarly, taking the final 18bases of the ORF, including the stop...will need to use a phosphatase to prevent re-circularization of the vector. You should treat your digested...Additionally, if your final product is going to be very large (>10kb) you might want to use electro-competent ...

  3. Western Blot

    Type
    Protocol
    ...high molecular weight proteins and higher percentage acrylamide when immunoblotting low molecular weight...and reagents from other vendors should produce similar outcomes when using these protocols. However, please... ideal lysis buffer will vary depending on the cellular location of the protein of interest. RIPA buffer...buffer will vary depending on the sample type and cellular location of the protein of interest. You may need...

  4. Stabilized Bacterial Promoters: Constant Gene Expression at any Copy Number

    Type
    Blog Post
    ...or other synthetic biology circuits. What is a molecular biologist to do? Let’s take a look at how plasmids...expression from the constitutive promoter did not. Similar levels of expression between a plasmid and the ...stabilized promoter, deoxychromoviridans titers were similar regardless of where the genes were expressed. What...

  5. Celebrating 15 Years of Scientific Sharing

    Type
    Blog Post
    ...worry about getting support from depositors. We regularly hear how much they see the value in depositing...) and even on 60 Minutes. We are blessed with a large community of scientists who support us by depositing... I checked we were ranked as the 21,806th most popular website in the world. I know, right? Upgrades to...

  6. RNA Extraction Without A Kit

    Type
    Blog Post
    ...Check out our molecular biology protocols Find protocol videos on our website Read our Molecular Biology Guide...many samples you may have, access to one or more large, expensive ultracentrifuge.  Figure 1: An ...

  7. Summer SciComm Series: Cognitive Load

    Type
    Blog Post
    ...bandwidth.  Jargon Learning, using, and retaining new vocabulary takes time and effort (otherwise it would be ...achieve!) Each field has its own framework: a particular way of organizing and connecting information ...experiences. If you’re hearing multiple people with similar misconceptions, it’s time to rework your strategy...

  8. Transferable Skills Guide: Problem Solving

    Type
    Blog Post
    ...Support Team here at Addgene, so troubleshooting molecular biology experiments was in the job description...fuzzy or ill-defined. In this article, I try to clarify what people mean when they ask about “problem solving...result of resolving the issue. Depending on the particular problem, you might even be expected to write ...

  9. The time and cost required to make a plasmid

    Type
    Blog Post
    ... repositories like Addgene provides large-scale savings. How large? Using Doench’s calculations and the...purchases can raise the cost anywhere from a few dollars to over a hundred. Of course, these were all 2018...

  10. Tagging Optogenetics and Chemogenetics Receptors: Fluorescent Proteins and Other Options

    Type
    Blog Post
    ...microscope showed us that, in monkey neurons, the large majority of the gold particles were in the cytoplasm...after it has been administered. Although we found a larger proportion on the membrane in mice neurons, most...the same cell, but not necessarily at the same subcellular compartment as the opsin or chemogenetic receptor...

  11. Hot Plasmids - March 2020 - base editors, GEVI, MoClo, and optogenetics

    Type
    Blog Post
    ...packaging capacity of 4-5 kb limits the delivery of larger transgenes, such as base editors which have a gene...efficiency, the split editors transduced at levels similar to that of a single AAV expressing GFP. The lab...cyanobacteria Article contributed by Andrew Hempstead The Modular Cloning (MoClo) system allows synthetic biologists...

  12. Pairing CombiGEM and CRISPR for Combinatorial Genetic Screening

    Type
    Blog Post
    ...CRISPR guide RNAs (gRNAs) for genome editing in large-scale has led to the rapid generation of genome-...combinatorially modify the genome, screen for a particular phenotype, and quickly profile the resultant ...Sciences of the University of Hong Kong. He is particularly interested in developing and applying new technologies...

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

    Type
    Blog Post
    ...sequence ranging from a single nucleotide change to large insertions like the addition of a fluorophore or...components. However, this sometimes requires intracellular injections into the cytoplasm or pronucleus ... albino mice ranging from 11% to 100%, a level similar to other approaches.  Nanoblades can also be used...

  14. CRISPR 101: Anti-CRISPR Proteins Switch Off CRISPR-Cas Systems

    Type
    Blog Post
    ...., 2020). Some Acr proteins are specific to a particular Cas protein while others can inhibit CRISPR enzymes...can increase the chance of off-target editing particularly after the majority of on-target editing has ...Cas9 Toxicity in Human Hematopoietic Stem Cells. Molecular Therapy - Methods & Clinical Development 9:390...

  15. Cpf1 Update: Comparison to Cas9 and NgAgo

    Type
    Blog Post
    .... Sticky-end mediated gene transfer would be particularly helpful for targeting non-dividing cells, which... are 3.7 kb and 3.9 kb in length, respectively, larger than SaCas9 but smaller than SpCas9. These nucleases...et al. suggest that AsCpf1’s off-target rate is similar to that of high fidelity Cas9s eSpCas9 and SpCas9...

  16. Plasmids 101: FLEx Vectors

    Type
    Blog Post
    ...point mutation or functional mutation in vivo: Similar to a conditional rescue experiment with a wildtype...sense (on) orientation in the presence of Cre. A similar strategy can be employed for "Cre-off" (i.e. your...for monitoring Cre-mediated recombination at the cellular level in the mouse. 2003 PubMed PMID: 12665802...

  17. Tips for Titering Your Lentiviral Preps

    Type
    Blog Post
    ... parallel, this approach should be considered. Similar to the p24 assay, titering via measurement of viral...serial dilutions of a viral prep. This method is particularly useful when vectors carrying fluorescent markers...In-Depth Fluoresence Titering Protocol Another popular approach utilizes the antibiotic resistance genes...

  18. Transferable Skills Guide: Managing a Team

    Type
    Blog Post
    ...yourself anymore. Personally I found this to be particularly challenging. As PhD students and postdocs we...there is not a clear deadline. Finally check-in regularly, make yourself available, and encourage questions...manager for advice if you’re not sure how to face a particular situation. But keep in mind that a good manager...

  19. Cloning Mammalian Cells with the Agarose Method

    Type
    Blog Post
    ...moist and hopefully aerated with a thin layer of molecular biology-grade agarose containing physiological... advantage of this technique is the potentially large number of single clones obtained (on occasion we...Maryland Medical School in Baltimore. She is particularly interested in signaling peptide maturation in...

  20. Truncated gRNAs for Regulating Gene Expression

    Type
    Blog Post
    ...Cas9 technology has revolutionized the fields of molecular biology and bioengineering, as it has facilitated...gRNAs Importantly, the use of truncated guides has largely been demonstrated to result in decreased mismatch...technologies. To date his group has develop several popular CRISPR-based tools for altering DNA sequence or...
Showing: 641 - 660 of 971 results