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We narrowed to 789 results for: mpl

Showing: 161 - 180 of 789 results

  1. IDP and your PI: A Roadmap for Career Planning and Personal Development

    Type
    Blog Post
    Published
    July 21, 2015, 2:30 p.m.
    ...credit for what we have accomplished. Starting with a recap of accomplishments, from learning new techniques...tangible research and career goals, and celebrate accomplishments. These techniques are laid out in Vincent et...DePace’s yearly planning meetings revolve around two simple worksheets, each of which is filled out separately...conversation. Goals and Planning Worksheet: Lists accomplishments, research/professional goals and feedback. ...including major project goals and effort needed to complete them. These documents dictate the subject matter...concrete plan in hand, trainees are motivated to complete their work rather than overwhelmed or confused...Extracurricular Activities for a Strong Science Career Path 7 Simple Strategies to Resolve Conflict With Difficult Supervisors...

  2. Tag Your Favorite Yeast Genes with Ease

    Type
    Blog Post
    Published
    Nov. 19, 2013, 2:37 p.m.
    ...desired tags have been integrated. Simply design your amplification primers with the desired targeting...cerevisiae and S. pombe. The protocol is surprisingly simple, requiring only a PCR product containing the modifying...markers. These are especially useful in protein complex studies where tagging of multiple protein products...for easy capture and detection of proteins and complexes, without the artifacts sometimes associated with...Formosa, at the University of Utah, has built a complete collection of yeast tagging modules with each ...ideal for tandem affinity purification of protein complexes. John Pringle and Jürg Bähler have deposited a...GST, or GFP). Longtine et al.(3) describe a complimentary set of plasmids for use in S. cerevisiae, with...

  3. Bioinformatics at Addgene

    Type
    Blog Post
    Published
    July 14, 2022, 1:15 p.m.
    ...needed to transform the raw sequencing data into a complete plasmid sequence that would be ready for our QC...containers are they are designed to ‘contain,’ in a complete package, all of the pieces of software that depend...can be easily shared with others without any complicated setup. We created, tested, and debugged each ...bioinformatics workflow in Nextflow Establishing a complete pipeline however, required more than just identifying...executed on Linux, we created our first process by simply copying and pasting pieces from our existing script...from the first process to the second process. For example, our bbduk process created trimmed FASTQ files ...sure we understood the concept of channels and implemented the input declaration correctly, the script block...

  4. An Inside Look into Addgene's Dev Team

    Type
    Blog Post
    Published
    Jan. 23, 2020, 2:45 p.m.
    ...approaches that may come up during discussion.  One example of agility at work is our process for working on...instructions on how to demonstrate the feature. For example, we recently reduced the number of steps involved...stories in that sprint relate to the theme. For example, we could have a shipping sprint and all the stories...get different perspectives on how things are implemented and from that, develop better software.  Maintenance...how you combine it with other ingredients.  For example, we use a third party library to handle label printing... compatible with the updated library. A recent example is updating our software from Python 2.7 to Python...analogy. Human languages evolve over time, for example from Ancient Greek to Modern Greek. Books written...

  5. How to Deposit Your Plasmids with Addgene

    Type
    Blog Post
    Published
    Feb. 27, 2024, 2:15 p.m.
    ...a peer-reviewed journal. To start this process, simply enter your preprint title or a general description...information directly into the file. Email the completed spreadsheet to [email protected] along with ...used to knockdown expression of mRNAs with complementarity to the shRNA sequence. Empty backbone A plasmid...plasmid data in the pages described below. You can complete these pages sequentially by entering the appropriate...any functional consequences of that change. For example, "Mutated Aspartate 123 to Lysine (D123K), lowers...box for backbones requested through Addgene. For example, you could enter "Feng Zhang (Addgene plasmid #52961...the links at the top of the page. Figure 11: A completed plasmid page for review before submission. Once...

  6. A Needle in a Base-Stack: Cas9 Structural Biology

    Type
    Blog Post
    Published
    June 4, 2024, 1:15 p.m.
    ...structurally resemble any other known proteins; they are completely unique to the CRISPR system (Nishimasu et al.... these regions, even if they are present in the sample. Other techniques, like electron microscopy and...conformation.   Fun Fact! Even though the Cas9:RNA complex is well described in the closed conformation, recent...electron microscopy studies have also found Cas9:RNA complexes in the open conformation (Cofsky et al., 2022)...guide RNA, prearranged for base pairing with a complementary DNA strand. These 10 nucleotides are the “seed...the PAM sequence and doesn’t mind if the PAM complementary strand is mismatched.   Figure 5: S. ... found a PAM site, Cas9 needs to check for complementarity with its guide RNA. But how to test DNA bases...

  7. Adeno-associated Viruses (AAVs) for Genome Editing

    Type
    Blog Post
    Published
    March 27, 2018, 1:32 p.m.
    ...one AAV and a repair template in second a second AAV to deliver their repair template to mouse zygotes and...system frequently over the past fifteen years. Some examples from their lab include using AAV to introduce ...homologous recombination with a homologous repair template. It is this application - the creation of sequence-specific...by using a targeted nuclease. Asuri et al, for example, used zinc fingers to increase the efficiency of...allowing a researcher to fill it with the repair template. Bok and Porteus 2017 recently showed that by ... as RNPs and delivering two halves of a repair template spread across two AAVs, it is possible sequentially...this technique, Bok and Porteus deliver a repair template that would take up ~6.5 kB if delivered in a single...

  8. 3 Tips to Improve HDR Efficiency for CRISPR Editing in Human Cells

    Type
    Blog Post
    Published
    Sept. 5, 2017, 1:58 p.m.
    ...the blocking-only templates will compete with the blocking/intended mutation templates resulting in cells...HDR), a distinct cellular DNA repair pathway. Accomplishing this usually involves simultaneously introducing...introducing a homologous DNA repair template, such as a sin­gle-stranded oligodeoxynucleotide (ssODN), which... – Edit the cells a second time using a repair template that corrects the blocking mutation and again ...that the cell was only using a part of the repair template and mutations further away from the cut site were...intended mutation site are available, a repair template containing blocking and intended mutations can...can be equally mixed with another ssODN template that contains the same CRISPR/Cas-blocking mutation but...

  9. CRISPR 101: Validating Your Genome Edit

    Type
    Blog Post
    Published
    Nov. 3, 2022, 12:15 p.m.
    ...trace file generated in Sanger sequencing and a simple algorithm to quantify editing efficiency by insertion...   How to perform a TIDE First, use PCR to amplify the target region of interest. Then Sanger sequence...unedited population and your Cas9-targeted cells. Simply upload both trace files to the TIDE tool online...flanking the editing site on either side of the amplicon. Based on the frequency of out-of-frame edits...have to screen to identify a knockout clone. For example, if your cell line is diploid (2N) and your out...Validating knock-in mutations You’ve introduced a donor template along with your gRNA and Cas9 combo and are hoping...deleted accordingly. Pro tip: make sure your PCR amplicon isn’t too large so that you can easily visualize...

  10. Multifunctional Neural Fibers: Modulation from Gut to Brain

    Type
    Blog Post
    Published
    Aug. 31, 2023, 1 p.m.
    ...interact during various behaviors. Either tissue samples were harvested after the behavior, or the animals...develop these fibers, the labs looked at previous implementation of optogenetic tools, calcium sensors, and ... be, as well as other features necessary for implantation in brain tissue. The gut, on the other hand...hand, was the wild west for fiber implants. Due to the soft nature of the intestines and the requirement...fibers could not be directly repurposed for gut implantation. The fibers they ended up generating were 10...relatively static). To operate both types of implanted fibers simultaneously, a mini-Bluetooth system...also record tissue temperature proximal to the implanted fibers. As important as its capabilities are the...

  11. R Bodies: Membrane-Rupturing Microscopic Tools

    Type
    Blog Post
    Published
    April 14, 2016, 2:30 p.m.
    ...understanding these complex processes can’t be overstated, we can also learn a lot from Nature’s simpler solutions...engineer it to suit our purposes? In search of simple biological machines R bodies can repeatedly ...These questions can be addressed using R bodies: simple, force-generating protein machines that can extend...conformational changes. These tiny movements are then amplified tens or hundreds of thousands of times by propagating...could be used in a variety of applications. For example, you could use them to lyse cells (and surrounding...

  12. Resume Writing for Non-academic Science Careers

    Type
    Blog Post
    Published
    Dec. 18, 2014, 4:06 p.m.
    ... could be meaningful for your potential employer. For example, my experience in science outreach and writing...have merely two pages to convince your potential employer that you are the right candidate for the job. ...and tailor the resume for the job posting. For example, if the job asks for experience in “statistical...French-speaking is an asset,” mention “speaks French.” Employers usually don't spend more than 20-30 seconds on...skills you developed through your research? Don't simply write about the research you did. Emphasize the...necessary in order for you to do your research. For example, did you help put together a grant application?... into a communications role instead after she completed her MSc in Experimental Medicine. She is the Community...

  13. Technique: Probe Phage Genomes for Host Binding Proteins

    Type
    Blog Post
    Published
    Sept. 29, 2016, 2:30 p.m.
    ... whole genome amplification (by a mechanism known as multiple displacement amplification) even in the ...faces as it approaches a host cell surface are complex polysaccharide structures, many of which are easily... DNA from host cell nucleases (4). To further complicate matters, even when sequenced, phage genomes tend...paper for more details on the assay, including examples of RBPs successfully identified using this method... for cell-binding abilities (6). This can be accomplished using a restriction enzyme with a short recognition...that is refractory to enzymatic manipulation, amplifying phage DNA with phi29 polymerase prior to fragmentation...others in liquid culture, dilute and plate the complete library of transformants for single colonies (...

  14. Plasmids 101: Screening Strategies Used in Plasmid Cloning

    Type
    Blog Post
    Published
    Sept. 15, 2020, 1:15 p.m.
    ...𝛽-galactosidase. Complementing this mutation by expressing this deleted portion, for example on a plasmid,...fewer colonies after your experiments. As a common example, a selection will leave you with the colonies that...plasmid contains your insert of interest or not. For example, choose enzymes that only cut once on either side...you will just see one band. These are only a few examples of how you can use restriction digest to screen... by using primers that are vector specific but amplify a potential insert. If you’re using insert-specific...no insert in the plasmid, you won’t be able to amplify anything. Like any experiment, you’ll want the ...get started, you will first need a primer that complements your plasmid sequence. Start with a backbone-...

  15. Plasmids 101: Yeast Vectors

    Type
    Blog Post
    Published
    Feb. 25, 2014, 7:11 p.m.
    ...cerevisiae yes - This can complement ura4- S. pombe, but the complementation is weak. yes   LYS2 L-lysine...vectors? Yeast are eukaryotes and thus contain complex internal cell structures similar to those of plants...mechanism of homologous recombination to facilitate simple gene replacement/mutation. Yeast expression plasmids... including a copy of a functional gene which complements the host’s auxotrophy. When grown on media NOT...LEU2 L-leucine no S. cerevisiae yes - This can complement leu1- S. pombe, but multiple copies are required...drug selection methods feasible in yeast as a complement or alternative to using auxotrophic markers. ...antibiotic resistance cassettes is most commonly employed, with the KanMX being the most prevalent due to...

  16. Kiran Musunuru on the Newest TALEN Genome-Editing System

    Type
    Blog Post
    Published
    Dec. 10, 2013, 3:13 p.m.
    ... the cell with a mutant template and fooling the cell into using that template for repair. By using these...the two ends back together or using a homologous template provided by the sister chromosome. As cells take...especially well-suited for use in stem cells. (See the complete protocol at StemBook). Addgene: How are you now...diseases. We are knocking out genes that have been implicated in disease or, in cases where particular variants...disease process. In the Cell Stem Cell paper, for example, we generated hepatocytes to examine the effect...much all investigators. Our kit makes it extremely simple to design TALENs. It only takes two to three days...less than a month's time. Addgene: What are the implications of this kit in terms of understanding human ...

  17. RANbodies: Reporter Nanobody Fusions

    Type
    Blog Post
    Published
    April 10, 2018, 12:56 p.m.
    ...detected with tyramide signal amplification. Tyramide signal amplification occurs when HRP converts a fluorescently...used at a time. Samples can be sequentially stained with two HRP RANbodies if the sample is stripped. HRP-RANbodies...Tyramide signal amplification of HRP Staining cultured cells and tissue sections Amplifying weak signals...process that’s difficult for individual labs to complete. The nanobody based RANbody platform from the ... lower background and better penetration of the sample due to the smaller size of RANbodies compared to...

  18. Pairing CombiGEM and CRISPR for Combinatorial Genetic Screening

    Type
    Blog Post
    Published
    April 12, 2016, 2:30 p.m.
    ...but what if we had a simple tool that allowed us to rapidly decode the complexity? The CombiGEM-CRISPR...was contributed by guest blogger Alan Wong. The complexity of biological systems can hinder our attempts...of providing an easy-to-use tool to analyze the complex combinatorial genetic networks underlying your ...experiments. CombiGEM-CRISPR: The marriage of two simple platforms CRISPR has revolutionized how we decode...mapping the combinatorial genetics that underlie complex regulation in biological systems. The CombiGEM ...new technologies to interrogate and understand complex biological systems. He can be contacted at aslw...transcriptional activation by an engineered CRISPR-Cas9 complex." Nature (2014). PubMed PMID: 25494202. PubMed ...

  19. Plasmids 101: Using Transposons in the Lab

    Type
    Blog Post
    Published
    July 6, 2021, 1:28 p.m.
    ...QCascade complex surveys the genome and binds its target DNA sequence. The transposase complex binds the...really depends on where a transposon lands. For example, in bacteria, transposons can move antibiotic resistance...is left behind. When the transposon-transposase complex finds its target site, it integrates, which produces...TEs You can think of autonomous transposons as “complete transposons” since they encode the protein they...selecting a transposon for a mutagenesis screen. For example, piggyBac and Tol2 are best for screening promoters...DNA-targeting module with the crRNA, 3) a transposase complex made up by three transposase proteins (TnsA, TnsB...

  20. Savvy Advocates Needed to Navigate a Scientific Enterprise in Flux

    Type
    Blog Post
    Published
    Sept. 13, 2016, 2:30 p.m.
    ... opportunities for us all.  For example, implementation and compliance for the FLSA changes, including...turn. The first is the passing and upcoming implementation of the Federal Labor Standards Act (FLSA) which...remain mixed in their opinions on the subject – for example, Yale graduate students filed for union elections...union dues. Time will tell the extent of the implications of this ruling and whether graduate students...finding money in already tight lab budgets and even simply being able to categorize all postdocs at institutions...challenging, but will be aided by the development and implementation of new strategies designed to cope with current...
Showing: 161 - 180 of 789 results