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We narrowed to 563 results for: cat.3

Showing: 481 - 500 of 563 results

  1. Bioinformatics at Addgene

    Type
    Blog Post
    ... the subsequent bbmerge process.     Fig. 3: How we get bbdone with our bbduk process.    ... from depositing laboratories and associated publications, but our quality control (QC) standards have...such as 96-well microplates, and thanks to our dedicated product and development teams, frequent updates...can be easily shared with others without any complicated setup. We created, tested, and debugged each ...

  2. Antibiotic Resistance: An Old Solution but a New Problem

    Type
    Blog Post
    ...Government (2016). 2. CDC Information Pages on Antibiotics 3. WHO Antimicrobial Resistance Fact Sheet  Additional...Microbes (bacteria included) are very skilled at the delicate art of survival. Existing virtually ubiquitously...of intentions. The take-home? Always take your medication as prescribed. How do we know which bacteria ... is passionate about health equity, science communication, and public health.     References 1. O'Neill...

  3. Light Sheet Fluorescence Microscopy

    Type
    Blog Post
    ...Frank Bradke laboratories later developed the 3DISCO (3-dimensional imaging of solvent cleared organs) method...of LSFM microscopes (3,4). After this initial application, Stelzer’s group described the single-plane or...brightness with increasing depth caused by light scattering and absorption.  Finally, the photomultipliers...

  4. CRISPR Library Amplification

    Type
    Protocol
    ...recover, set up overnight growth (Estimated time 2-3 hours) Transformation should be performed at the end...Day 2: Harvest cells and purify DNA (Estimated time 3-4 hours) Cells should be harvested first thing in ...µL micropipette tips in a -20 °C freezer. Aliquot 3 mL SOC into each of four 14 mL Vented Falcon Tubes...cuvette and add to 14 mL vented Falcon Tube containing 3 mL SOC. Repeat for each of the 25 µl aliquots of cell...Vented Falcon Tubes should contain a total of 5 mL (3 mL SOC + 2 mL transformed Endura from two separate... Protocols CRISPR Library Amplification CRISPR Library Amplification You may also like... Pooled libraries...quantities of library for experimental applications. Repeated amplifications should be avoided as best as possible...

  5. Lab to Office Culture Shock

    Type
    Blog Post
    ...school. The transition has been a bit rough for her (3 weeks in and there's a big office vs. lab culture ...involves a few key features that can be difficult to replicate in other work settings: The sense of being the... might include collaborating, managing, and communicating. After years of focusing on my technical skills...

  6. Transferable Skills Guide: Career Planning Resources

    Type
    Blog Post
    ...www.businessinsider.com/how-to-get-the-most-out-of-job-fairs-2014-3 https://www.forbes.com/sites/neilpatel/2015/05/21/... conferences are held every year in different locations all over the world. But simply attending the conference...work. Addgene’s very own Joanne Kamens, a strong advocate for career planning, has written a great blog ...

  7. Troubleshooting and Optimizing a Western Blot

    Type
    Blog Post
    ...the membrane is cut so each lane is its own piece (3). After incubation, blocking, and washing, the membrane...protein concentration decreases).     Figure 3: Testing the antibody’s dynamic range. A) The protein...check its subcellular location and solubility. If it’s membrane-bound or located in a subcellular compartment...nuclear- or DNA-binding proteins, you may need to sonicate your lysates in order to release the proteins ...will simply load an extra ladder or technical replicate (ie, leftover sample) to fill the relevant wells...membrane into the psi membrane, that’s a good indication that the p membrane pores are too large.  Transfer...likely technical. White circles, for example, could indicate that you had air trapped between the membrane ...

  8. AAV Purification by Iodixanol Gradient Ultracentrifugation

    Type
    Protocol
    ... of 25% iodixanol step 5 mL of 40% iodixanol step 3 mL of 60% iodixanol step Carefully add up to 5 mL ... tubes. Repeat for each QuickSeal tube. The first 3 mL collected corresponds to the 60% phase and can ...from the 40% phase contain the purified AAV. Option #3 Puncture the QuickSeal tube slightly below the 60–...Link opens in a new window) copyright (2006). Figure 3: Example of an SDS-PAGE gel of 10 consecutive gradient.... The arrow indicates the 60–40% interface. The vertical black line indicates the location of the purified... Protocols AAV Purification by Iodixanol Gradient Ultracentrifugation AAV Purification by Iodixanol Gradient...isomolar density gradient medium suitable for virus purification and isolation of cells, organelles, lipoproteins...

  9. Negotiating Work and Life: How to Find the Joy

    Type
    Blog Post
    ...100th birthday, as your mom will never forgive you… #3 Honor the work day and keep it holy Keep a regular...other activities. Take care of yourself. Take a vacation alone with your partner – no kids. Get a massage...

  11. Designing Your Chalk Talk for the Academic Job Interview

    Type
    Blog Post
    ...acceptable. Aims should be mechanistic, doable by 1-3 people in 1-5 years, and contribute to resolving the...anticipated outcome, and payoff for each aim. Learn to communicate your story with only the most salient details...

  12. Supporting Reproducibility with a Connected ELN

    Type
    Blog Post
    ...connectivity to other tools used in research; and (3) connectivity to open source data repositories. To... both accept deposit of data associated with publications into one of five general purpose open source...

  13. General Transfection

    Type
    Protocol
    ...transfected using 1:1, 1:2, 1:3 and 1:6 µg of pRosetta :µg of PEI. The 1:2 and 1:3 ratios provided high transfection...transfection - Remove media, replace with fresh media Day 3 or more (am): Observe fluorescence, harvest cells,...high levels of virus. HEK293T cells should be split 3 times a week: Monday: Plate 1x10 6 cells in a 75 cm...negative E. coli strain such as NEB stable. Dilute 1:3 (µg DNA:µg PEI) in 500 µL total of OptiPro SFM (per...of 1 mg/mL PEI (μL) 1:1 18.9 18.9 1:2 18.9 37.8 1:3 18.9 56.7 1:4 18.9 75.6 1:5 18.9 94.5 1:6 18.9 113.4...inhibit transfection; therefore, plasmid DNA purification should include an endotoxin removal step. For...

  14. Neurodegeneration Plasmid Collection

    Type
    Collection
    ...Spinocerebellar ataxia 22 Bernard Weinstein 21238 PKC gamma CAT PRKCG HA CMV Spinocerebellar ataxia 21 Bernard Weinstein...TLS 2: hTLS.pCDNA1 FUS CMV ALS David Ron 21829 TLS 3: TLS.pRSET.C FUS His T7 ALS David Ron 21830 TLS 4:... APEX2 CMV Parkinson's Mark Cookson 165107 pGEX4T-3-6AOptParkin PRKN GST tac Parkinson's Arthur Haas 166671...dementia (FTD) Thomas Kukar 176922 pcDNA3.1(+)_Granulin 3+linker4 GRN Flag, Strep CMV Frontotemporal dementia...dementia (FTD) Thomas Kukar 176923 pcDNA3.1(+)_Granulin 3-no linker GRN Flag, Strep CMV Frontotemporal dementia... FUS His T7 ALS Nicolas Fawzi 226605 FUS LC 4S->A#3 FUS His T7 ALS Nicolas Fawzi 226606 FUS LC 8Y->S FUS... HA CMV ALS Catherine Tomasetto 104450 pEGFPC1-hVAP-B KD/MD VAPB GFP, HA CMV ALS Catherine Tomasetto 104465...

  15. Lentivirus Production

    Type
    Protocol
    ...total DNA to μg PEI ratios of 1:1, 1:2, 1:3 and 1:6. The 1:2 and 1:3 total DNA:PEI μg ratios provided high...post-transfection. Remove media, replace with fresh media. Day 3-4 (am): Harvest virus Equipment Class II, Type A2 ...obtaining high viral titer. 293T cells should be split 3 times a week: Monday: Plate 1×10 6 cells in a T75 ...enough PEI such that the ratio of μg DNA:μg PEI is 1:3 (1000 μL total per 10 cm dish). Using transfer plasmid...DNA μL of 1 mg/mL PEI 1:1 18.9 18.9 1:2 18.9 37.8 1:3 18.9 56.7 1:4 18.9 75.6 1:5 18.9 94.5 1:6 18.9 113.4...lentivirus can be used for a variety of downstream applications such as stable-cell line generation. Workflow...without calcium or magnesium, Corning 21-040-CV (cations can affect the attachment of adherent cells) 0.45...

  16. CRISPR Guide

    Type
    Guide
    ...EQR variant 3' NGAG SpCas9 VQR variant 3' NGAN or NGNG xCas9 3' NG, GAA, or GAT SpCas9-NG 3' NG Staphylococcus...SP); SpCas9 3' NGG SpCas9 D1135E variant 3' NGG (reduced NAG binding) SpCas9 VRER variant 3' NGCG SpCas9...jejuni (CJ) 3' NNNNRYAC Neisseria meningitidis (NM) 3' NNNNGATT Streptococcus thermophilus (ST) 3' NNAGAAW...the table above, 3' and 5' indicate on which end of targeted sequence the PAM is located. The majority of...continue to anneal to the target DNA in a 3′ to 5′ direction. The location of any potential mismatches matters...double-strand break (DSB) within the target DNA, located ∼3–4 nucleotides upstream of the PAM sequence. The...Each library typically contains ∼3–6 gRNAs per gene to ensure modification of every target gene, so CRISPR...

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

    Type
    Blog Post
    ...pCas into a non-temperature-sensitive replicon, and 3) added the sacB gene for counterselection to yield...far, we’ve used the MUCICAT system in two ways: In our initial paper, we used MUCICAT to optimize the gene...industrial strain. Not only is MUCICAT better, it is faster. Using MUCICAT only needs 2 newly constructed...integration using CRISPR-associated transposases” or MUCICAT. Here, we’ll describe each of these CRISPR systems...providing users with a complete editing process. The MUCICAT system While the above systems are great for experiments...chromosomal integration (ex: genome editing), our MUCICAT system which is based on the CRISPR-associated ...unstable and lack precise control over copy number. MUCICAT system contains 4 plasmids: pDonor, pTnsABC, pQCascade...

  18. AAV Titers: Where Do They Come From And What Do They Mean?

    Type
    Blog Post
    ..." Gene therapy 6.7 (1999). PubMed PMID: 10455443. 3. Hindson, Benjamin J., et al. "High-throughput droplet...the other to guess a category by listing off things that fall into that category? Okay, let’s play. I’...’ll list the examples and you try to guess the category: ELISA...qPCR...Digital droplet PCR...DNA dot ...concentration of viral genomes (by qPCR or other DNA quantification methods - see below), since each viral particle...value, physical titers don’t necessarily give any indication of the infectious titer. The infectious titer...affected by many experimental factors. To list some complicating factors for quantifying AAV: qPCR efficiency...optimized our assay in a few ways:  Absolute quantification by qPCR requires generating a standard curve...

  19. CRISPR 101: Which Cas9 Do I Choose for My CRISPR Experiment?

    Type
    Blog Post
    ...types can be found on Addgene’s website. Fig. 3: Cas9 repressors include dCas9 alone and a dCas9-KRAB... than ever to make precise, targeted genome modifications. Cas9 has been modified to enable researchers...instead.  Choosing a Cas9 for site-specific modifications Site-specific deletions and knockouts Some experiments...mutation at a specific site e.g., deleting the catalytic residue of a protein or knocking out a very small...flexibility as to where your sgRNA needs to be located and there may not be a NGG PAM site anywhere to... Beyond SpCas9.” Single base pair edits Is the location and type of edit you need to introduce highly ...Perhaps you need to introduce a single nucleotide modification at a defined amino acid to mimic a patient mutation...

  20. Changing Labor Laws Bring Increased Postdoc Wages

    Type
    Blog Post
    ... level should now be automatically adjusted every 3 years. In May 2016, Secretary of Labor Thomas Perez...Labor’s general guide to the effect on higher education, you can see that those in teaching roles (such...frankly quite surprising. Consortia of higher education institutions submitted letters to the Department...centrally administered, which leads to many complications, including the inability of some institutions...paid. Institutional salary policies are one indicatator of how salaries may changes due to the FLSA. ...of the postdoctoral workforce. Based on our communications with and research on these institutions, we...to the new level:  We are still waiting for an indication of what (most of) Harvard, Johns Hopkins, (most...
Showing: 481 - 500 of 563 results