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We narrowed to 911 results for: IRE

Showing: 441 - 460 of 911 results

  1. To Codon Optimize or Not: That is the Question

    Type
    Blog Post
    ... the ideal codon for every amino acid across an entire polypeptide. Many researchers have created codon-optimization...necessary for proper protein folding (Stadler and Fire, 2011). Therefore, codon optimization may disrupt...1987) The codon adaptation index-a measure of directional synonymous codon usage bias, and its potential...doi.org/10.1016/s0165-0270(02)00362-x Stadler M, Fire A (2011) Wobble base-pairing slows in vivo translation...

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

    Type
    Blog Post
    ...DNA strain which results in two DSBs that can be repaired by NHEJ restoring mCherry expression. Cells can...populations. To develop this method the lab was inspired by work that used an integrated GFP reporter that...fluorescent protein) upon CRISPR/Cas9 homology directed repair (HDR) (Glasser et al., 2016). For TREE ...different reporter proteins. Protein translation requires the start codon of AUG immediately downstream ...

  3. Plasmids 101: Plasmid Incompatibility

    Type
    Blog Post
    ...  Antisense RNAs encoded on the plasmid either indirectly inhibit replication through inhibiting translation...translation of replication machinery proteins, or directly inhibit replication by binding to the origin of...sequences within the origin of replication and are required for replication in plasmids that contain them ...Kamruzzaman M, Shoma S, Thomas CM, Partridge SR, Iredell JR (2017) Plasmid interference for curing antibiotic...

  5. Designing Your Chalk Talk for the Academic Job Interview

    Type
    Blog Post
    ...This post was contributed by Erik Snapp, the Director of Student and Postdoctoral Programs at the Janelia...single model. The first approach can lead to new directions, while the second approach could terminate with...think realistically about the time and resources required to feasibly accomplish the aims and clarify what... and Structural Biology, and now serves as the Director of Student and Postdoctoral Programs at the Janelia...

  6. Transferable Skills Guide: Creativity

    Type
    Blog Post
    ...skill developed during PhD training. Creativity is required to develop new projects and test novel approaches... all the funding in the world? What if you could hire more people? What would you do with unlimited time...over half of which, when implemented, impacted the entire company. It is important to note that the person...product team is an incredibly innovative group. Our entire company benefits from their policy! Perfectionism...

  7. Plasmids 101: Shuttle Vectors

    Type
    Blog Post
    ...helps increase cellular uptake, but this approach requires compatible cell lines that express viral machinery...ribosome. One resistance gene on your vector might require two different selection molecules! It all depends...option is to use a different method of selection entirely. Auxotrophic selection relies on the host organism...organism being unable to synthesize a nutrient required for growth. Before delivery of the vector, the deficient...

  8. Plasmids 101: Codon usage bias

    Type
    Blog Post
    ... and therefore may need fewer resources for the entire translation process. For example, during high growth...inappropriate locations or fail to make it through the entire transcript resulting in the production of nonfunctional...that their codons are more appropriate for the desired expression host. This is known as “codon optimization...Addgene now have full sequence data, we recommend directly analyzing gene sequences for codon optimization...

  9. Four Ways to Package Transgenes That Exceed the Size Limit of Adeno-associated Virus

    Type
    Blog Post
    ...AAV donor template for CRISPR-mediated homology directed repairs (HDR). Split AAV vector approaches Split...trans-splicing (62%) vector. 4. Sequential homology directed repairs Overview: The sequential homologous recombination... donor templates using two sequential homology directed repairs (HDR) with CRISPR/Cas9. Donor A contains...M, Colella P, Garcia-Hoyos M, Gibbs D, Kim SR, Maguire A, Rex TS, Di Vicino U, Cutillo L, Sparrow JR, ...

  10. Antibodies 101: Single Chain Fragment Variables (scFvs)

    Type
    Blog Post
    ...be expressed in bacteria, while mAbs generally require mammalian expression systems. scFvs are also small...to avoid animal use by using semi-synthetic or entirely synthetic VL and VH sequences. Phage display also...for the scFv (Gaciarz and Ruddock., 2017), but requires an oxidizing environment to form. When expressed... age-related macular degeneration (AMD) since direction injection to the eye results in high drug concentrations...

  11. Plasmids 101: Expressing shRNAs for RNA Interference

    Type
    Blog Post
    ...composition of the RISC varies, but at the very minimum requires an Argonaute protein. This Argonaute protein is...shRNAs using plasmids The design of shRNA plasmids requires three important steps: Designing the shRNA sequence...This is much shorter than miRNA plasmids, which require additional flanking regions comprised of upstream...35(9), 872–878. https://doi.org/10.1038/nbt.3947 Fire, A., Xu, S., Montgomery, M. K., Kostas, S. A., Driver...

  12. Viral Vectors 101: Integrase-Deficient Lentiviral Vectors

    Type
    Blog Post
    ...Ciuffi, 2012). One family of viruses, retroviridae, require integration for their life cycle to continue. They...IN (integrase). These viral packaging genes are required for the production of lentiviral particles.  The... keep in mind is that titering these IDLVs may require a slightly different protocol than regular lentiviral... (2008). Insertional mutagenesis combined with acquired somatic mutations causes leukemogenesis following...

  13. Hot Biosensors 2022: Year-End Roundup

    Type
    Blog Post
    ...variety of behaviors, opening up new research directions that were inaccessible with earlier tools. They... in localization to the cell membrane and not a direct change in the fluorescent properties, pYtags are...variations and heat transfer at the subcellular level requires a probe with fast kinetics and high signal/noise...temperature indicator B-gTEMP designed to meet those requirements (Lu et al. 2022). By fusing two bright fluorescent...

  14. Multiple Plasmids at a Low Price: Kits from Addgene

    Type
    Blog Post
    ...bypasses the 5′ mRNA cap required for eukaryotic systems and instead directly engages the ribosome for... protocol and are compatible with single-end or paired-end read flow cells. FREQ-Seq is an open source... The kit contains all the variant PUF segments required to bind any 8 base RNA sequence. The segments ...

  15. Antibodies 101: The Basics of Western Blotting

    Type
    Blog Post
    ...overnight.  In contrast, a semi-dry transfer only requires enough buffer to saturate the sandwich and typically...without any transfer buffer. These systems, however, require you to purchase expensive system-specific sandwiches...on the antibodies and labels being used and may require some trial and error (Yang et al., 2012).  Choosing... they provide qualitative measurements. If you require precise quantification of a protein, consider using...

  16. Antibodies 101: Multiplex Immunofluorescence

    Type
    Blog Post
    ...secondary antibodies altogether and multiplex with direct IF, using primary antibodies that are conjugated...However, the primary-secondary workflow (a.k.a. indirect IF) is more flexible and efficient, since you ...primaries for whichever targets you like. Plus, indirect IF amplifies the signal since multiple secondary...design. Like selecting an antibody, some of the requirements are similar to standard IF. Here, we’ll walk...

  17. Plasmids 101: Dimers and Multimers

    Type
    Blog Post
    ... on as a dimer, depicted on the right, with the entire monomeric sequence duplicated in tandem, resulting... with large inserts (>7 kb)(Berg et al., 1989), direct repeats (Ribeiro et al., 2009), and at high copy...frequency plasmid recombination mediated by 28 bp direct repeats. Mol Biotechnol. 2008 Nov;40(3):252-60....Summers D. Timing, self-control and a sense of direction are the secrets of multicopy plasmid stability...

  18. Finding Your Perfect Job After University

    Type
    Blog Post
    ... degree. I loved parasitology. However, when I enquired about roles in this field, most jobs needed at...understood why parasitology roles in the field require experience. Working in industry: Microbiology ...redundancies can be made within the company; new staff hired in anticipation of the contract suddenly have less...institutes. The lab at the University of Queensland required a scientist for a 6 month project to validate ...

  19. Mycoplasma Contamination: Where Does It Come From and How to Prevent It

    Type
    Blog Post
    ... to stain all DNA. Some amount of expertise is required in this process because result interpretation ...serum to be used for subculturing. Avoid using expired products or reagents that have been stored for ...cross-contamination. Make sure you keep all the required equipment and reagents inside the laminar airflow... Biotechnology.       References Degeling MH, Maguire CA, Bovenberg MSS, Tannous BA (2012) Sensitive ...

  20. Typing CRISPR Systems

    Type
    Blog Post
    ...systems has led to extreme sequence divergence that requires a multi-pronged analysis to sort effectively. ...efficiency when conducting edits using homology-directed repair. There are 10 subtypes of type V, named...select RNA transcripts, or be engineered to conduct direct RNA editing. There are four subtypes, A–D, with...surface-level function of each type could lead you to new directions in your research. We all have our preferences...
Showing: 441 - 460 of 911 results