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We narrowed to 385 results for: abo.5

Showing: 181 - 200 of 385 results

  1. The PAM Requirement and Expanding CRISPR Beyond SpCas9

    Type
    Blog Post
    Published
    Aug. 20, 2020, 2:30 p.m.
    ... Protospacer Adjacent Motif (PAM sequence). The 3-5 nucleotide PAM sequence serves as a binding signal... Acidaminococcus sp. BV3L6 Cas12a, AsCpf1, uses a 5'-TTN PAM which makes it easier to target AT-rich genomes...Resources on the Addgene Blog  Read more about Cpf1 Read more about SaCas9 and AAV Check out our CRISPR Featured... and VRER) recognize novel PAM sequences (shown above). The VQR, EQR, and VRER Cas9 variants are capable...example, Cas9 from Staphylococcus aureus (SaCas9) is about 1 kilobase smaller than SpCas9, so it can be packaged...

  2. 3 Tips to Improve HDR Efficiency for CRISPR Editing in Human Cells

    Type
    Blog Post
    Published
    Sept. 5, 2017, 1:58 p.m.
    ...also takes into account that HDR is rare, usually 2-5% of the edited cells in our hands, these combined ...heterozygous event, the cut should ideally be around 5 to 20 bp away from the cut site. Lastly, if a heterozygous...mutations When thinking further about the distance relationship described above, we realized that, in addition...efficiency already drops by half and after only about 30 bp, it is no longer feasible to incorporate mutations...Additional Resources on the Addgene Blog Learn about High Fidelity Cas9 Variants How to Design Your gRNA...

  3. Deep Mutational Scanning with One Pot Saturation Mutagenesis

    Type
    Blog Post
    Published
    Feb. 22, 2017, 3:30 p.m.
    ...MgCl2, 4) increased concentration of dNTP, and/or 5) the addition of MnCl2. Disadvantages of this method...PMID: 25167058. PubMed Central PMCID: PMC4412028. 5. Araya, Carlos L., and Douglas M. Fowler. "Deep mutational... all desired codon changes (step 1 in the image above). A second universal primer is used to synthesize...first followed by the top strand. Both produced about the same number of positive clones. See Supplementary... of DNA degraded in the first digest (step 1 in above diagram). Remember that the BbvCI enzymes nick the...mutagenesis with the same protocol. What do you think about one pot saturation mutagenesis? How would you use...

  4. Controlling for Off-target Effects with a New Genome-wide CRISPR Screen Design

    Type
    Blog Post
    Published
    Sept. 13, 2018, 1:55 p.m.
    ... mismatch off-targets were only toxic if they had 5+ off-target sites. Results from the screen also reproduced...Pubmed PMID: 23394947. Pubmed Central PMCID: 4299491. 5. Bassik, Michael C., et al. "A systematic mammalian...following unanswered questions about CRISPR screen design. Unanswered questions about genome-wide CRISPR/Cas9...Morgens et al drew some generalizable conclusions about off-target activity, demonstrate safe-targeting ...

  5. Overcoming the AAV Size Limitation for CRISPR Delivery

    Type
    Blog Post
    Published
    Sept. 16, 2020, 1 p.m.
    ...cells using compact expression cassettes. Sci Rep 5: . https://doi.org/10.1038/srep10777  Garneau JE, ...:97–110 . https://doi.org/10.1038/s41551-019-0501-5  Liu J-J, Orlova N, Oakes BL, Ma E, Spinner HB, Baney...Addgene Blog Learn more about Cas9 Variants and the Expanded CRISPR Toolbox Read about CRISPR enzyme Cpf1 ... and specific? The two AAV strategies described above showed successful target modification, indicating...

  6. Antibodies 101: Reading a Flow Plot

    Type
    Blog Post
    Published
    Feb. 6, 2024, 3:13 p.m.
    ...suggest using the imaginary quadrant to help (Figure 5). Even though this real-world data is far messier ...different populations on a plot.      Figure 5: The gating strategy from Barlow-Anacker et al., 2017...series of one- or two-marker plots, like in Figure 3 above, until the population of interest has been successfully...  Of course, there are many more things to learn about when diving into flow cytometry data. There’s different... ways to use FSC and SSC, as briefly alluded to above. There’s many variations in how one can gate, select...

  7. Important Considerations in Optogenetics Behavioral Experiments

    Type
    Blog Post
    Published
    Oct. 1, 2015, 2:30 p.m.
    ...acquisition and retrieval of contextual fear memories [5].  In a modified behavioral paradigm, the authors ...primate cortex." Frontiers in systems neuroscience 5 (2011). PubMed PMID: 21811444. PubMed Central PMCID...of the important considerations you should think about when developing optogenetics behavioral experiments...scientific research is that the field benefits from collaboration and the exchange of ideas. The rapid spread ...versatility and standardization of genetic tools in the laboratory mouse, most optogenetics is performed in this...

  8. Synthetic Biology, Artificial Chromosomes, iGEM & More - Interview with Tom Ellis

    Type
    Blog Post
    Published
    Nov. 29, 2017, 2 p.m.
    ...microbial cooperation, and antibiotic production. Chapter 5: The future of Synthetic Biology. Wide adoption of...synthetic yeast chromosome. Listen to learn more about Dr. Ellis, synthetic biology, synthetic chromosomes...Additional Resources from the Addgene Blog Learn about the Synthetic Biology Education Nonprofit BioBuilder...BioBuilder Plasmids 101: Golden Gate Cloning Learn About Working at a SynBio Startup in Our Interview with...

  9. Four Base Editing Reporters to Monitor and Enrich Editing in Real-time

    Type
    Blog Post
    Published
    July 7, 2020, 1:15 p.m.
    ...APOBEC-Cas9n-UGI targets its preferred trinucleotide motifs (5’-TCA-to-TUA) found within the 43 base-pair mCherry...Genome Editing. Molecular Therapy - Nucleic Acids 5:e334 . https://doi.org/10.1038/mtna.2016.48 Katti ...for base-editing activity The reporters mentioned above all rely on GFP fluorescence and cell sorting using... Additional resources on the Addgene blog Read about other CRISPR reporters Browse our CRISPR blog posts...

  10. The Addgene Antibody Data Hub

    Type
    Blog Post
    Published
    Sept. 12, 2024, 1:15 p.m.
    ...conditions described above, and a short description of how the antibody performed.  Figure 5: Results for the...

  11. AAV Titers: Where Do They Come From And What Do They Mean?

    Type
    Blog Post
    Published
    Nov. 15, 2017, 3 p.m.
    ... real-time PCR. Nat Methods. 2013 Oct;10(10):1003-5. PubMed PMID: 22035192. PubMed Central PMCID: PMC3216358...PMID: 20486768. PubMed Central PMCID: PMC2957240. 5.Taylor, Sean C., Genevieve Laperriere, and Hugo Germain... Addgene.org Find All Available AAV Learn More about Virus Production Find Viral Vector Protocols ...

  12. New Acoustic Reporter Genes: Ultrasound Imaging of Gene Expression

    Type
    Blog Post
    Published
    Jan. 24, 2023, 2:15 p.m.
    ... cells in Lennox LB (10 g/L tryptone, 5 g/L yeast extract, and 5 g/L NaCl) or on LB-agar plates containing...you enjoy playing with our genes. Have fun!    About the authors: Marjorie Buss is a PhD student studying...

  13. 6 Tips for Analyzing and Troubleshooting Sanger Sequencing Results

    Type
    Blog Post
    Published
    June 26, 2014, 2:23 p.m.
    ... least 50bp upstream of the sequence of interest. 5. Use a silica spin column for purification of the ...and troubleshooting your DNA sequencing results Above (top right corner of this post) is an example of...

  14. Save Time and Money by Making Your Own Competent Cells

    Type
    Blog Post
    Published
    Jan. 28, 2014, 3:10 p.m.
    ...rather than the ZymoBrothTM, so it costs us less than 5 cents per transformation. This protocol has worked...own competent cells using the protocols provided above, then you can find the subsequent transformation...

  15. Quick, Versatile Plant Transgenesis with GreenGate Plasmids

    Type
    Blog Post
    Published
    Jan. 23, 2014, 3:23 p.m.
    ...High Throughput Capability." PLoS ONE. 3(11): e3647 (5 November 2008).  ...protein,” he says. “There is nothing you can do about it but be gentle. If it no longer works, just trash...

  16. Five Popular Model Organisms, Part 2

    Type
    Blog Post
    Published
    Sept. 29, 2020, 1:15 p.m.
    ... Human Genetic Disease. Cells Tissues Organs, 205(5–6), 303–313. https://doi.org/10.1159/000490898 Gibbs.... J. (2009). Rats! Disease Models & Mechanisms, 2(5–6), 206–210. https://doi.org/10.1242/dmm.002733 Idalia... more about model organisms in the first Five Popular Model Organisms blog post Read more about plasmids...characteristics that make them ideal for studying in a laboratory environment. We’ve covered five model organisms...have been specifically developed to thrive in laboratory settings. One of its most attractive features...minutes, they are perfect for molecular cloning and metabolic engineering.  The E. coli genome has an extensive...address many areas of microbial research like metabolism. Transport, biofilms, and much more. Because ...

  17. March for Science

    Type
    Blog Post
    Published
    April 21, 2017, 2:30 p.m.
    ...consumption in comparison to 37% of the general public (5, 6). Similarly 90% of active research scientists believe...activity while much less of the general public agrees (5, 6). Many of the new administration’s actions, including... with a testable hypothesis - an educated guess about how some part of the world functions. Experiments...governments exist in reality and science is how we learn about reality. Formally, a government can be defined ... More concretely, scientists gather information about reality informing people and their governments on...people to rally for science, others are concerned about possible unintended consequences of a march. Three...Democrats more often than Republicans, the March is not about Democrats or Republicans (16). This is a Pro-Science...

  18. Plasmids 101: Common Lab E. coli Strains

    Type
    Blog Post
    Published
    Nov. 7, 2014, 2:56 p.m.
    ...rB- mB-) recA13 leuB6 ara-14 proA2 lacY1 galK2 xyl-5 mtl-1 rpsL20(SmR) glnV44 λ- JM109   General cloning...hsdS20(rB-, mB-) ara-14 galK2 lacY1 proA2 rpsL20 xyl-5 leu mtl-1 Stbl4   Electrocompetent cells for cloning...thiamine metabolism Requires exogenous thiamine for growth ara disruption of arabinose metabolism pathway...little prokaryotic minions that carried out the labor-intensive process of replicating your new plasmid... Phage resistance gal Mutation in galactose metabolism pathway Cells cannot grow on galactose only ...repressed more   LacZ β-galactosidase activity abolished   lacY Lactose permease inactivated, lactose...plasmid stability recBCD Exonuclease V activity abolished Increased plasmid stability, reduced recombination...

  19. Stem Cell Models for Disease & Open Science: Interview with Darrell Kotton

    Type
    Blog Post
    Published
    July 7, 2017, 1:32 p.m.
    ...Regenerative Medicine accelerates research. Chapter 5: Research currently under way in the Kotton Lab. One...promote scientific sharing. Listen to learn more about Darrell, his research, and his focus on open science...Medicine and in the Department of Pathology and Laboratory Medicine. He attends in the Medical Intensive...NIH-funded Principle Investigator in the Kotton Laboratory and is the founding Director of the Center for...Podcast.     Additional Resources from Addgene Learn about Retinal Gene Therapy from Connie Cepko Find Plasmids...Plasmids from the Kotton Lab Learn more about Stem Cells ...

  20. Fluorescent Proteins 101: When GFP lets you down

    Type
    Blog Post
    Published
    Aug. 23, 2018, 12:05 p.m.
    ...deleting residues. Only about 10 amino acids can be deleted from the C-terminus and about 5 from the N-terminus... 153.7 (2013): 1602-1611. PubMed PMID: 23768684.  5. Thorn, Kurt. "Genetically encoded fluorescent tags...cylinder with a length of 4.2 nm and a diameter of about 2.4 nm (Hink et al., 2000). The complete beta-barrel...based on cofactor binding proteins as described above. The smallest tags are peptides that are engineered...
Showing: 181 - 200 of 385 results