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We narrowed to 397 results for: abo.4

Showing: 61 - 80 of 397 results

  1. What's the Best Way to Elute and Store Your Plasmid DNA?

    Type
    Blog Post
    ...periods of time, especially at room temperature or at 4°C. Storing DNA: temperature and longevity You should...be stable for years. Plasmid DNA can be stable at 4°C or even room temperature for a short period, and...to purify DNA without a kit Read our blog posts about plasmid cloning Browse our Plasmids 101 blog posts...

  2. The Effect of COVID-19 on Liu Lab Plasmid Requests From Addgene Data

    Type
    Blog Post
    ...had a tremendous impact on our world -- as of June 4, 2020, COVID-19 has caused an estimated total of 380,000...deaths worldwide (statistics provided by Google on 6/4/20). Many countries entered lockdown for weeks to ...during weekends and significant holidays. Figure 4: Plasmid request trajectories suggest an exponential...in his laboratory. Learn more here. Additional resources on the Addgene blog Learn more about plasmid...

  3. AAVs CREATed for Gene Delivery to the CNS and PNS

    Type
    Blog Post
    ... for more than making pretty pictures (see figure 4). It also enables single-cell anatomical study and...49 +/- 7% 36 +/- 8% 37 +/- 10% AAV.PHP.eB 69 +/- 4% 55 +/- 7% 76 +/- 5% Table 1: Transduction efficiency...induction of multi-color labeling of neurons (figure 4). These capsids can serve as powerful cell-type-restricted...or the PNS, respectively.  Read on to learn more about these two new AAV capsids and how they can be useful...Important considerations when using AAVs Learn more about using AAV as a tool for gene expression in mammals...

  4. MXS Chaining

    Type
    Blog Post
    ...Membranes 3 mCherry 587nm/ 610 nm human β-Actin Actin 4 Citrine 516nm/ 529nm human α-Tubulin Tubulin Repeatable...methods. 2009. May;6(5):343–345. Pubmed PMID: 19363495. 4. Josse J, Kaiser AD, Kornberg A. Enzymatic synthesis...Approaches in Mammalian Systems. PLoS One. 2015 Apr 24;10(4):e0124958. PubMed PMID: 25909630. PubMed Central PMCID...Resources on the Addgene Blog Read about Gibson Assembly Learn About Other Plasmid Cloning Techniques Plasmids...

  5. Tips for Screening with Yeast Two Hybrid Systems

    Type
    Blog Post
    ...POI’s) are fused to TAs and DBDs (see Figure 2a) (4). These fusions are then co-expressed in a yeast strain...bacterial LexA DBD and the lacZ reporter gene (3, 4), there are now many variations, including many that...repressor. Cell. 1985; 43:729-736. PubMed PMID: 3907859. 4. Fields, S., Song, O. A novel genetic system to detect... Plasmids Read our Blog Post About Yeast Vectors Read our Blog Post About Tagging Yeast Genes ...

  6. Plasmids 101: Gateway Cloning

    Type
    Blog Post
    ...an attL-containing entry vector.   Figure 4: Method B to create an entry clone: TOPO cloning the...vectors at once in the same tube. You can clone up to 4 DNA fragments, in a specific order and orientation...Technology: Advantages and Drawbacks. Clon Transgen 4:138. doi:10.4172/2168-9849.1000138 2. Hartley JL. ... expression systems. Keep reading to learn more about the Gateway cloning method and its advantages. An... on the Addgene Blog Try Out FastCloning Learn About Methylation and Restriction Enzymes Use Colony PCR...

  7. Avoiding the Dark Side of Fluorescent Protein Fusions with mOX FPs

    Type
    Blog Post
    ...function in the environment where it will localize [4]. Most investigators are surprised to learn that FPs...biology."Current Protocols in Cell Biology (2005): 21-4. Pubmed PMID: 18228466. Pubmed Central PMCID: PMC2875081...Pubmed PMID: 6526011. Pubmed Central PMCID: PMC557822. 4. Costantini, Lindsey M., and Erik Lee Snapp. "Fluorescent...moxFPs over standard FPs for the reasons listed above. In addition, because the moxFPs are highly monomeric...Medicine. Her doctoral research was conducted in the laboratory of Erik Lee Snapp and focused on optimizing fluorescent...

  8. Lambda Red: A Homologous Recombination-based Technique for Genetic Engineering

    Type
    Blog Post
    ...alternative approach is to flank the desired change with 4-5 silent changes in the wobble codons, i.e. make changes... changes to the third base pair of the adjacent 4-5 codons that alter the nucleotide sequence but not ...defective prophage 2) from a plasmid, 3) from mini-λ or 4) from the lambda red phage itself (each of these will...2016. doi:10.1128/ecosalplus. PubMed PMID: 27223821. 4. Sharan SK, Thomason LC, Kuznetsov SG, Court DL. Recombineering...currently a research scientist in the Department of Laboratory Medicine at the University of Washington. She... Additional Resources on the Addgene Blog Read about Homology Directed Repair Use Knockout/Knockin Plasmids...

  9. Cre-lox system

    Type
    Collection
    ...pAAV-CA-FLEX Cre activates gene of interest, using 4 mutated lox sites AAV Uchida Cre Reporters and Tools...Recombination Between loxP Sites. J Mol Biol. Aug 25;1504(4):467-86. doi:10.1016/0022-2836(81)90375-2. PubMed ...Learn about the Cre-Lox system and how it can be used for genetic rearrangements and recombination. Use...

  10. Some Like it Hot: Thermostable GeoCas9

    Type
    Blog Post
    ... SpyCas had similar levels of editing. See figure 4 in Harrington et al for more examples of GeoCas9’s...with reaction buffer for 8 hours at either 37 °C or 4 °C. Substrate was then added and cleavage product ..., see the table below and read on to learn more about GeoCas9.   SpCas9 GeoCas9 Size 1368 AA 1087...vivo. What aspect of GeoCas9 are you most excited about? Thermostability? Longevity in human plasma? Something... Additional Resources on the Addgene Blog Read about CRISPR Methods for Bacterial Genome Engineering   ...

  11. Sleeping Beauty Awakens for Genome Engineering

    Type
    Blog Post
    ...transposon; the plasmid is then degraded by the cell.4.) The transposase creates a double strand break in...stable transgenic cell lines. Biotechnol J. 2015 Feb 4. doi: 10.1002/biot.201400821. PubMed. 2. Mates L, ...of DNA that can move around in a genome. In a laboratory setting, transposons can be used to both introduce...

  12. Live and Let Dye: Self-Labeling Protein Tags

    Type
    Blog Post
    ... A flexible chemical tag. Nature Methods, 2(4), Article 4. https://doi.org/10.1038/nmeth749 Strebinger...HaloTags and SNAP-tags. Cell Chemical Biology, 26(4), 584-592.e6. https://doi.org/10.1016/j.chembiol.2019.01.003...Proximity-Induced Reactivity. ACS Chemical Biology, 4(7), 547–556. https://doi.org/10.1021/cb900062k Gautier...but it’s real! In this post, we’re going to talk about the power and flexibility of self-labeling protein...ready-to-use without any cloning at all! As mentioned above, several variants of these tags have been developed...Illuminating Choices blog post for more discussion about choosing a dye or ligand to use for your experiment...

  13. Filming Science Videos in the Age of Social Distancing

    Type
    Blog Post
    ...and chat with me remotely via an audio chat (Fig. 4).  Learn: This works great for on-camera presentations...back to producing protocol videos?   Figure 4: The remote directing setup, complete with wireless...camera control, and extra hand sanitizer.    Cycle 4: Venturing into the Lab Design: The lab is the beating... Production Specialist, I’m constantly thinking about ways to improve our productions to deliver the best...four months after I started with Addgene, I was about to have a check-in with my boss Jennifer. I had ... would be a couple months. Jennifer and I spent about an hour that morning scrambling. Everything we had...someone to be comfortable on camera while thinking about things like lighting, camera angle, and shot composition...

  14. CRISPR-based Genome Editing Systems For Editing in E. coli

    Type
    Blog Post
    ... successfully tested the pEcCas/pEcgRNA system in 4 E. coli K-12 strains, 2 E. coli B strains, 1 E. coli...control over copy number. MUCICAT system contains 4 plasmids: pDonor, pTnsABC, pQCascade and pCutamp.  ... constructed plasmids (pDonor and pQCascade) with 4 rounds of transformation to complete the N-acetylglucosamine...complete editing process. The MUCICAT system While the above systems are great for experiments that require only...serve to interrupt genes (or gene clusters) in metabolic engineering. So far, we’ve used the MUCICAT system...when compared to the industrial strain. In our metabolic engineering case study of N-acetylglucosamine ...Multiple induction systems to choose from. Ready for metabolic engineering with improved cargo dedicated to gene...

  15. Deep Mutational Scanning with One Pot Saturation Mutagenesis

    Type
    Blog Post
    ...extension time, 3) increased concentration of MgCl2, 4) increased concentration of dNTP, and/or 5) the addition...glycosylase (UDG) and Exonuclease III (ExoIII) (step 4). UDG recognizes uracils that are in DNA and ExoIII...Methods 13.11 (2016): 928-30. PubMed PMID: 27723752.  4. Fowler, Douglas M., Jason J. Stephany, and Stanley... 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...

  16. Tips for CRISPR Gene Editing in Mice

    Type
    Blog Post
    ...fluorescence intensity was classified into four groups (4; brighter than control, 3; same as control (Cetn1/...you can hold off on genotyping until the pups are 2-4 weeks old and use the ear tag method (creating different...differentiation 56.1 (2014): 122-129. PubMed PMID: 24372541. 4. Wang, Haoyi, et al. "One-step generation of mice ...the pX330 plasmids used in the validation assay (above) and it is just as effective (Mashiko et al., 2014...Engineering with CRISPR Validate Your Genome Edit Learn about Non Homologous End Joining and Homology Directed...

  17. Getting the Most from Your Lentiviral Transduction

    Type
    Blog Post
    ...storing for less than a day, lentivirus can be kept at 4°C. For long-term storage, viral preps should be divided...expression profiles.” Biotechniques 35.4 (2003). 812-4. Pubmed PMID: 14579747. 3. Drexler, H.G., Uphoff, ...Cytotechnology 39.2 (2002). 75-90. Pubmed PMID: 19003295. 4. Uphoff, C.C., Drexler, H.G. “Detection of mycoplasma...Lentiviruses are a powerful laboratory tool often employed to establish cell lines that stably express...Get Tips for Titering Your Lentivirus Learn All About Viral Vector Elements Learn How to Do Genome-Wide...

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

    Type
    Blog Post
    ...manipulate prokaryotic cells? Especially, if you only have 4 hours and 35 students! After a course coordination... the format I decided on was to have two teams of 4 people face each other on a single chosen clinical...manipulation more hands on While teaching courses about gene therapy and genetic manipulation, I and other...with some other professors, I went home thinking about how to solve this problem. I didn’t want to face...therapeutic strategies work. Find Addgene Blog posts about other educational tools Playing BUILD YOUR PLASMID...winner? Ultimately, society should be! This is a collaborative game, as it should be in real scientific practice...practice transferable skills like team building, collaborative critical thinking, and communication among others...

  19. An Introduction to Adenovirus

    Type
    Blog Post
    ...adenovirus using native viral promoters. Mol Ther, 7(4), 526–534. https:/doi.org/10.1016/S1525-0016(03)00023... human adenovirus infections. Am J Epidemiol, 117(4), 455–466. https://doi.org/10.1093/oxfordjournals....release of adenovirus from infected cells. J Virol, 70(4), 2296. https://doi.org/10.1128/JVI.70.4.2296-2306.1996...this introduction gives a sense of what AdVs are about – and we hope that it inspires further use of existing...

  20. CRISPR 101: Cytosine and Adenine Base Editors

    Type
    Blog Post
    ... positions 4-7. ABEs 6.3, 7.8, and 7.9 display slightly wider editing windows of position 4-9, although...with a 14% indel rate, while ABE7.10 modified only 4/12 off-targets with a frequency of 1.2%. Although ...editing strategies are there? CBEs and ABEs can induce 4 out of 12 possible base-to-base changes. The other...cytidine deaminase fusions. Nature Biotechnology, 35(4), 371–376. https://doi.org/10.1038/nbt.3803 Koblan...Homology-Directed Repair Learn about base editing for plants Read about four base editor reporters Additional... The BE3 system increased editing frequency to above 30% for a variety of targets in human cell lines... Liu lab created BE4max and AncBE4max mentioned above, they also made an adenine base editor with improved...
Showing: 61 - 80 of 397 results