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  1. Addgene's Tips for Plasmid Quality Control

    Type
    Blog Post
    ... 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...
  2. The CRISPR Software Matchmaker: A New Tool for Choosing the Best CRISPR Software for Your Needs

    Type
    Blog Post
    ... 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...
  3. Sensing Neuronal Dopamine

    Type
    Blog Post
    ...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...
  4. CRISPR 101: Drug discovery and CRISPR

    Type
    Blog Post
    ...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...
  5. Understanding GPCR Signaling Using TRUPATH

    Type
    Blog Post
    ...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...
  6. All in a Twist: dsRNA

    Type
    Blog Post
    ... 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...
  7. Addgene's Top 10 Blog Posts of 2015

    Type
    Blog Post
    ...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...
  8. Custom CRISPR Screens & the Green Listed Software

    Type
    Blog Post
    ...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 ...
  9. Using CRISPR/Cas9 to Edit Disease Out of the Genome

    Type
    Blog Post
    ...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...
  10. Plasmids 101: Mammalian Vectors

    Type
    Blog Post
    ... 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...
  11. Plasmids 101: Methylation and Restriction Enzymes

    Type
    Blog Post
    ...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 ...
  12. 3 Challenges in Plant Synthetic Biology

    Type
    Blog Post
    ...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...
  13. Plasmids 101: Sequence and Ligation Independent Cloning (SLIC)

    Type
    Blog Post
    ..., 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′ ...
  14. Visualizing Protein Turnover In Situ

    Type
    Blog Post
    ...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...
  15. Plasmids 101: Cre-lox

    Type
    Blog Post
    ..., 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...
  16. How to Prepare for an Industry Interview

    Type
    Blog Post
    ...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...
Showing: 301 - 320 of 439 results