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We narrowed to 824 results for: EED

Showing: 301 - 320 of 824 results

  1. Plasmids 101: Gateway Cloning

    Type
    Blog Post
    ... given set of resources, time, and experimental needs. Since its invention in the late 1990s, Gateway ...easily select for the expression clone. You will also need to use a E. coli strain sensitive to CcdB (e.g. ...clones can be used with a large variety of plasmids. Speed The Gateway system enables the generation of the...set up the BP and LR reactions in the same tube, speeding up the cloning of the attB-PCR products directly...

  2. Changing Labor Laws Bring Increased Postdoc Wages

    Type
    Blog Post
    ...issued a Notice of Proposed Rulemaking, soliciting feedback by Sept 4th.  They proposed raising the salary...for Dec 1 2016. In order to be exempted from the need to pay employees overtime, salaries must be above... of $23,660. Any postdocs on such a salary would need to have their salary raised 101% if hours are not...make gradual adjustments towards this new level. Indeed some institutions did this. It doubtless took a...

  3. Viral Vectors 101: Pseudotyping

    Type
    Blog Post
    ...To deliver genes using lentiviral vectors, you need an envelope protein on the virus’s surface and a ...Because rabies dG can’t infect neurons on its own, it needs to be pseudotyped. This takes advantage of the EnvA-TVA...pseudotyping that could be broad or specific to suit your needs. Check out some of these review articles to find...https://doi.org/10.2174/1566523054546224 DePolo NJ, Reed JD, Sheridan PL, Townsend K, Sauter SL, Jolly DJ...

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

    Type
    Blog Post
    ...updated pEcCas/pEcgRNA system, and confirmed that indeed the gRNA-pMB1 had a slightly higher leaky expression...MUCICAT better, it is faster. Using MUCICAT only needs 2 newly constructed plasmids (pDonor and pQCascade... strain library, our pEcCas/pEcgRNA system would need to construct 13 plasmids with 14 rounds of transformation...of these systems a try, we’d love to hear your feedback!   Thank you to our guest bloggers! Qi Li obtained...

  5. Five Popular Model Organisms, Part 2

    Type
    Blog Post
    ...organism; they are relatively simple to maintain and breed and they have large, easily modifiable embryos, ...frog research is The National Xenopus Resource, a breeding facility for X. laevis and X. tropicalis, the ...development, a large number of offspring (produces many seeds), self fertilizing, a small yet highly characterized...Wang, S. H., & Jiang, J. P. (2016). Research proceedings on amphibian model organisms. Dong wu xue yan...

  6. Antibodies 101: Introduction to Immunofluorescence

    Type
    Blog Post
    ...reduce non-specific binding of antibodies.  You may need to optimize the type of block used, the concentration...which will bind to your target molecule. You may need to optimize the concentration and incubation time...antibody and is conjugated to a fluorophore. You may need to optimize the concentration and incubation time...antibody seems and what sort of protocol you will need to use. (To learn more about antibody validation...

  7. Plasmids 101: Expressing shRNAs for RNA Interference

    Type
    Blog Post
    ...mature sequence in a plasmid and call it a day. You need to include both sides of the complementary RNA duplex...hairpin loop sequence (Figure 2A). Since you don’t need additional flanking sequences, shRNA plasmids are...issue with many gene silencing methods and just needs to be taken into account when designing experiments...of study for improving these tools. Next time you need to knock down a gene, maybe you’ll consider using...

  8. March for Science

    Type
    Blog Post
    ...scientists as well as other citizens. Thus civilization proceeds in an informed fashion. For science to be useful...representative in the House of Representatives all you need to know is your area code. Senators: To find contact...contact information for your senators all you need to do is select your state. State Legislatures: To find...Trends in Federal R&D NJ.com: Rush Holt: Lawmakers need to use a scientific approach to formulating views...

  9. Site Directed Mutagenesis by PCR

    Type
    Blog Post
    ...undesired mutations through the PCR process, you need a high-fidelity polymerase. There are many high ...high fidelty polymerases on the market; you need one with 5’->3’ polymerase activity (for amplification),...potentially truncate the template). Importantly, you also need a DNA polymerase that produces blunt-ended PCR products...DNA duplex stability from the base sequence." Proceedings of the National Academy of Sciences 83.11 (1986...

  10. Evolution of Lab Techniques

    Type
    Blog Post
    ...fluorometric-based detection. This increased sequencing speed ultimately leading to what are generally considered...became the workhorses of molecular biology." Proceedings of the National Academy of Sciences 102.17 (2005...restriction endonuclease of Hemophilus influenzae." Proceedings of the National Academy of Sciences 68.12 (1971...sequencing with chain-terminating inhibitors." Proceedings of the National Academy of Sciences 74.12 (1977...

  11. Plasmids 101: Gibson Assembly and Other Long-Homology Based Cloning Methods

    Type
    Blog Post
    ...choice as shown in the figure below. To start, you need to have DNA fragments with regions of homology at...step without scars as depicted below. First, you need to design primers to amplify the two fragments while...is typically fairly high, so there is usually no need to screen a large amount of colonies. Excluding ...fragments. CRISPR cutting was used to circumvent the need to PCR amplify the fragments of the chromosome, ...

  12. Evolution of Brainbow: Using Cre-lox for Multicolor Labeling of Neurons

    Type
    Blog Post
    ...previous blog post, Plasmids 101: Cre-lox, if you need a quick primer on how Cre-lox recombination works...provided neuroscientists with the vast array of colors needed to mark individual neurons, the system also suffered...distinct cell populations. To decrease the animal breeding necessary to produce animals with Brainbow transgenes...provides spatial and temporal control without the need for germline modification, and enables Brainbow ...

  13. Easi-CRISPR: Generating Knock-In and Conditional Mouse Models

    Type
    Blog Post
    ... would breed them many times, hoping to produce a mouse with the desired edit. You’d also need to remove...remove the selection marker, likely by breeding to a mouse line with a recombinase. If it turned out your...design a knock-in allele at a given location, you’ll need to design only a single guide and your donor ssDNA...

  14. CRISPR 101: Off-Target Effects

    Type
    Blog Post
    ...off-target effects, you still need to do your due diligence to ensure they are indeed a non-issue. Above we mentioned...You only successfully generate 1 null clone and proceed with it. In this case, the likelihood of having...factors. Hopefully now you have all the resources you need to determine what your risk level is, how to lower...

  15. Technique: Probe Phage Genomes for Host Binding Proteins

    Type
    Blog Post
    ... proteins with host-binding activity without the need for genome sequencing or annotation (5). In the ... shearing exist as well, such as sonication and needle shearing. Essentially, any method that results ...forceps. Several protein induction conditions may need to be tested, as with any protein purification, ...DNA fragments with live host cells Now you can proceed with incubating this membrane with your cells of...

  16. An “elegans” Approach to Better CRISPR/Cas9 Editing Efficiency

    Type
    Blog Post
    ...approaches rapidly enrich for editing events without the need for any selective marker to remain in the edited...events in a sea of unedited animals struck me as a “needle in the haystack” type of problem and led to me ...result in efficient editing. Repair oligos do not need to be PAGE purified, although doing so increases... that is mainly lost in F2 animals, removing the need to segregate away co-conversion markers. Expanding...

  17. A Practical Guide to Optimizing AAV DIO and FLEx Vector Expression

    Type
    Blog Post
    ... the optimal/minimal injection titer and volume needed to achieve adequate expression in your cells of...volumes to determine the optimal injection parameters needed to transduce your cells of interest. Using a constitutively... range of dilutions and/or injection volumes may need to be tested with each new vector or lot used.   ...methods section to include all of the variables that need to be accounted for (i.e. serotype, promoter, titer...

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

    Type
    Blog Post
    ...replication.  Once the recombineering system is no longer needed, bacteria can be “cured” by growing them at 42℃...not only allows for activation of the red genes needed for recombineering, but also leads to expression...the modification into a different background, if needed. This approach is best suited for modifying plasmids...Red-mediated recombinants in mismatch repair mutants. Proceedings of the National Academy of Sciences of the United...

  19. 22 Hot Plasmid Technologies from 2014

    Type
    Blog Post
    ...all-synthetic vector that included only the elements needed for function. The lab validated this vector, called...Wittbrodt might be just the set of tools that you need. The Golden GATEway cloning kit simplifies the cloning...require expensive peptides or small molecules and exceed a physiologically relevant exposure in intensity...engineering which drastically reduces the time and effort needed to insert new genes into bacteria. They designed... simplifying experiments when additional cross-breeding steps are undesired or infeasible. Finally, an...

  20. Find and Share AAV Data with Addgene's New AAV Data Hub

    Type
    Blog Post
    ...questions! How much virus should I inject? How long do I need to wait? What serotype and promoter should I use...serotypes, expression patterns, and more. If you have feedback or suggestions about the AAV Data Hub, we welcome...welcome you to share your thoughts through our Feedback Form.  Thank you for your participation in this ...
Showing: 301 - 320 of 824 results