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  1. DNA Purification Without a Kit

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    Blog Post
    ...also less accessible due to their expense and requirement for lab equipment. Plus they create plastic waste... discreet sizes (mini, midi, maxi, giga) which requires you to buy different kits for different purposes...nucleic acids are released by dipping the cellulose directly into a PCR buffer, where DNA or RNA can be amplified...This method takes less than 30 minutes and only requires incubating the columns with 1M phosphoric acid...
  2. Transferable Skills Guide: Identifying Your Transferable Skills

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    Blog Post
    ...description. You check off all the boxes - you have the desired degree, you have mastered the applications they...person. Similar reasoning may also apply to other acquired skills.” It is nice when other people point out... of a time you’ve lead your team in a defined direction in the past. Maybe there was a time you noticed...list all the skills that your current or past job requires day-to-day, and you’ll get a great snapshot of...
  3. Course-Based Undergraduate Research Experiences (CUREs)

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    Blog Post
    ...students published their work identifying new genes required for stem and progenitor cell maintenance in planarians...Kimball et al., 2020). Benefits and Challenges CUREs inspire students to take ownership of their work and become...Biology Department at Keene State College in New Hampshire. He mentors undergraduates conducting NIH- and...Pellettieri, J. 2020. The exon junction complex is required for stem and progenitor cell maintenance in planarians...
  4. Introducing Addgene's New Search - Find Plasmids More Easily For Your Research!

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    Blog Post
    ...content-aware suggestion dropdown list will let you jump directly to the type of information you are looking for...straightforward search is now even easier with a direct link to the plasmid page at the bottom of the dropdown... dropdown list: The search bar will suggest direct links to other types of searches including depositing...search for “Melina Fan” and you’ll be able to go directly to a page to see the plasmids deposited by our...
  5. AAVs CREATed for Gene Delivery to the CNS and PNS

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    Blog Post
    ...screen for IV deliverable AAVs? Typically AAVs are directly injected into the brain, but this tends to limit... (figure 4f) of the tTA inducer virus, but the desired sparsity of labeling is achieved with lose doses...-color labeling of neurons, because it doesn’t require the generation of a transgenic mouse. 2. Cell-... gene vectors for neurons, no transgenic mouse required (see figure 5 and supplemental figure 8 in Chan...
  6. Tips for Screening with Yeast Two Hybrid Systems

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    Blog Post
    ...binding-dependent expression of the HIS3 gene, which is required for histidine manufacture and therefore growth...of interest, or a defined cDNA library (e.g. the entire proteome of a particular cell type, or a subset...at the same time in order to try and access the entire interactome of a set of proteins at once.  However...gain access to the nucleus. To circumvent this requirement for transmembrane proteins, a split ubiquitin...
  7. The Importance of a Fun Workplace: Company Culture at Addgene

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    Blog Post
    ... Chen co-founded Addgene in 2004. They worked tirelessly to get Addgene from a concept to an up and running...previous years, this event was held at Executive Director Joanne Kamen’s home- a testament to the familial...it! 8. Organize If you’re doing an event that requires people to be in groups, organize your troops prior... this saying is especially true. If your event requires a lot of set up or cleanup, ask for volunteers...
  8. Transferable Skills Guide: Creativity

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    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...
  9. Rett Syndrome: A History of Research and Therapeutic Outlooks

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    ...Rett syndrome experience regression of previously acquired skills such as speech, coordination, purposeful...programs in biopharma. AveXis, co-founded by Kaspar, acquired by Novartis and rebranded to Novartis Gene Therapies...adeno-associated viral (AAV) vector. The editase repaired half of the mutant MECP2 RNA in each of three ...Samaco RC, Zoghbi HY (2011) Adult Neural Function Requires MeCP2. Science 333:186–186 . https://doi.org/10.1126...
  10. To Codon Optimize or Not: That is the Question

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

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    ...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 ...
  12. Plasmids 101: Plasmid Incompatibility

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    ...  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...
  13. Harnessing TnpB for Genome Editing: A Compact and Versatile Tool for Your Research

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    ...cleave both strands. Similar to how Cas proteins require a PAM (protospacer adjacent motif) near the target...a different TAM (5′-TTAT-3′). The unique TAM requirements expand the number of genomic sites that can ... and prime editing (Molla et al., 2021), which require fusing Cas proteins with additional enzymes or ...transformation into plant cells. Conclusion & Future Directions TnpB represents a new generation of compact, ...
  14. QC Sequencing Technologies at Addgene

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    ...strand is synthesized (Figure 1). Sanger sequencing requires four separate reactions, where a fraction of one...in ultra-long read mode), enough to sequence an entire plasmid in one read. Runs can be completed in anywhere...sequencing, as well as the ability to sequence an entire viral genome in one read. We also use Nanopore ...keeping up with the large amount of QC analysis required every week. They are constantly exploring ways...
  15. Plasmids 101: Shuttle Vectors

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    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...
  16. Designing Your Chalk Talk for the Academic Job Interview

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    ...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...
  17. Plasmids 101: Codon usage bias

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    ... 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...
  18. Hot Biosensors 2022: Year-End Roundup

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

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

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    ...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, ...
  21. Plasmids 101: Expressing shRNAs for RNA Interference

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    ...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...
  22. Viral Vectors 101: Integrase-Deficient Lentiviral Vectors

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

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    ...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 ...
  24. Antibodies 101: The Basics of Western Blotting

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    ...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...
  25. Antibodies 101: Multiplex Immunofluorescence

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    ...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...
  26. Viral Vectors 101: AAV Serotypes and Tissue Tropism

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    ...proteoglycan (HSPG)3 AAV receptor (AAVR; not strictly required)2 Human fibroblast growth factor receptor ...targeting of projection neurons and enhances astrocyte-directed transduction. Nature Communications, 14(1), 3792...Adeno-Associated Virus Serotype 4 (AAV4) and AAV5 Both Require Sialic Acid Binding for Hemagglutination and Efficient...adeno-associated virus type 5 to 2,3-linked sialic acid is required for gene transfer. The Journal of Biological Chemistry...
  27. Plasmids 101: Dimers and Multimers

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    ... 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...
  28. Finding Your Perfect Job After University

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

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    ... 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 ...
Showing: 421 - 450 of 912 results