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Showing: 181 - 200 of 948 results

  1. Getting the Most from Your Lentiviral Transduction

    Type
    Blog Post
    ... express a gene of interest. While the general approach for using lentivirus, infect and select, seems...mycoplasma can be extremely hard to detect with levels reaching 108 cells per ml before the media becomes cloudy... cell lines, ELISA, fluorometry, or PCR-based approaches to detect even low levels of contamination. For...Uphoff et. al. outline a do it yourself PCR-based approach complete with freely available positive controls...benefit of the multiple harvest strategy is that each day you are supplying your producer cells with fresh...envelope. One drawback of the multiple harvest approach is that it tends to result in a higher volume ... your lentiviral prep!) and infect. While this approach is a great start to a successful infection, one...

  2. Oh, The Places You Can Go: Careers in Science Communication - Product Development

    Type
    Blog Post
    ... was weird like that,” says Caitlin. A love of teaching also runs through Caitlin’s blood. “I come from...faculty members, I helped grad students who were teaching for the first time. I walk[ed them] through their...them advice, [and helped] proctor exams.” These teaching experiences led Caitlin back to being a student... signed up for a 10 week class at Iowa called “Teaching Your Research.” This course has participants take...you know about it [your area of research], and teaches you how to design a course, how to design good ...ground up.”  Science writing and editing These teaching experiences definitely weighed in Caitlin’s post-grad...but in the end, writing won out. “I considered teaching at a liberal arts school but I’ve always enjoyed...

  3. Antibodies 101: Normalization and Loading Controls for Western Blots

    Type
    Blog Post
    ...sample of your protein of interest (POI). This approach is reliable, but it’s overkill in many cases. ...normalized to the loading control: the intensity of each POI band is divided by the intensity of the loading...Figure 2: Normalization with loading controls. Under each lane, the fraction of the POI band over the loading...detected with an antibodies, just like the POI. This approach is so common that we’ve used it for our cartoon..., 2008; Gilda & Gomes, 2013). Depending on the approach you choose, your loading control measurement will...on the same gel or membrane (depending on your approach) as your final blot. Quantifying the entire lane...targets. Convenient! Comparison of loading control approaches Cartoons are nice and all, but we’ll leave you...

  4. Transferable Skills Guide: Conflict Resolution

    Type
    Blog Post
    ...that many of my old suitemates stopped talking to each other if I was not there to arbitrate for them. ...sell them. Types of Conflict Before explaining my approach to conflict resolution, it’s important to recognize...occurs when two people cannot agree or get along with each other, regardless of the situation. If left unchecked... of conflict can lead to people disagreeing with each other just for the sake of disagreeing. It can be...people or teams who normally get along well with each other disagree about a specific decision, direction...responsibilities. Knowing which aspects of customer support each team “owns” is critical for day-to-day operations...everyone walk away from a conflict happy, feeling they each got enough of what they wanted. However, a poorly...

  5. Antibodies 101: Introduction to Immunofluorescence

    Type
    Blog Post
    ...immunofluorescence There are a couple different approaches to be familiar with when talking about IF, direct...antibody targets the antigen of interest but is not attached to a fluorescent molecule, instead a secondary...BioRender.com   Indirect IF is a much more common approach than direct IF, so we will focus on this technique...why generating fluorophore-conjugated versions for each primary antibody could get out of hand. Instead,...can also require some additional optimization as each antibody performs differently and some antibodies.... There are multiple methods for fixing samples, each with its own pros and cons. The method you use will...immunofluorescence procedure with key considerations about each step. Image created with BioRender.com   The...

  6. Multiplexed Overexpression of Regulatory Factors (MORF) collection

    Type
    Blog Post
    ... was delivered to each cell, future applications could deliver multiple TFs to each cell, allowing for...The Zhang lab generated a comprehensive library of each unique TF open reading frame (ORF), barcoded for...target cells as a pooled library, which will amplify each TF to determine how its presence influences cell...atlas of directed differentiation To understand how each human TF can contribute to differentiation, the ...embryonic stem cells with the viral TF library, with each cell receiving only 1 TF expression construct. Cells...screening mechanisms, although the unique barcodes for each TF help streamline identification. While the Zhang...and/or TF dysregulation. A targeted TF screening approach can not only lead to a better understanding of...

  7. Affinity Purification of Recombinant Antibodies with Protein A or Protein G

    Type
    Protocol
    ...Starting Wipe down all pipettes and equipment with 10% bleach prior to use. Warm the affinity column, binding...gently pour off the ethanol storage buffer. Tightly attach LabMate PD10 Buffer Reservoirs to the top of the...columns, refer to the manufacturer’s instructions. Attach the Gravitrap column to a clamp on a clamp stand...s to mix. Determine the protein concentration of each fraction on the NanoDrop Spectrophotometer using... 3x , discarding buffer from the collection tube each time. Place column in a new 50 mL LoBind collection...NanoDrop Spectrophotometer to see if the sample has reached the desired concentration. If the sample concentration...until the volume of the sample is reduced enough to reach the desired concentration. Pro-Tip Periodically ...

  8. Antibodies 101: ChIP

    Type
    Blog Post
    ...Capturing), if you ask me.  Each of the three steps need to be optimized for each specific sample type. And...including epigenetic modifications. But it's not an approach for the faint of heart!  The Process ChIP can... your protein:DNA complexes will remain bound to each other until you are ready to separate them. Most...Most proteins are not likely to remain attached to the DNA throughout the sample processing and therefore...sample may need more or less cross-linking to firmly attach the proteins to the DNA, depending on how tightly... idea to add "patience and an extra-methodical approach" to the top of your protocols when embarking on... Rachel Leeson...

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

    Type
    Blog Post
    ...sequences (loxp and lox2272) and can not recombine with each other. When Cre is introduced to the system it binds...optimal/minimal injection titer and volume needed to achieve adequate expression in your cells of interest.  ...injection titers and volumes based on the serotype Since each serotype also exhibits distinct patterns of spread...waiting periods and/or higher injection titers to achieve adequate expression. Conducting a time course experiment...start an experiment before adequate expression is reached you may obtain a false negative or conclude the...to minimize off-target expression while still achieving adequate expression in your cells of interest..../or injection volumes may need to be tested with each new vector or lot used.   Following this stepwise...

  10. Mouse Modeling, Part 2: Breeding and Crossing Mice

    Type
    Blog Post
    ...knockin or knockout mice. While these techniques are each useful for introducing one modification into the...engineering should be used to separately introduce each mutation independently into different mice, and ...have a combination of the mutations engineered into each of their parents. Here’s how to cross mice, using... scheme should start with parental mice that are each homozygous for one of the two mutations that you...Genotyping involves getting a small piece of tissue from each mouse (typically via either a blood draw, tail clip...in handy here! Crossing your F1 heterozygotes to each other will generate approximately 1 in 16 mice that...gametes that contain a mix of genetic material from each of the parents. If both of the mutations that you...

  11. How-to: 5 Steps to a Great Panel Discussion

    Type
    Blog Post
    ...organizers and moderator should have a well-planned approach to making the event interesting and engaging. ...an organizer) should set up a 30 minute call with each panelist to get to know their style and to get their...discovering interesting stories, riffs or opinions that each panelist can contribute. The moderator should identify...information and remind them about the event as it approaches. Room set-up and logistics To make a really...formality and distance that prevents interaction with each other and the audience. Put the panel on a stage...panelists in a slight semi-circle (so they can see each other) on comfortable chairs (stools if you have... better. Have a good microphone system ready for each panelist if the room needs it (lapel mics are best...

  12. Cloning Mammalian Cells with the Agarose Method

    Type
    Blog Post
    ...transfection efficiency is not known, this is easily achieved by splitting transfected cells into 3 plates at...about 200 ul of culture medium containing serum in each well. A 200 ul pipettor is set to 200 ul, wiped ...ml sterile tube (for 60 mm dishes only 1.5 ml of each are used). This solution is capped, swirled and ...additional growth medium is added to about halfway up each well. Tips for using the agarose cloning method...the cell type, we sometimes add a drop of serum to each well in the 48-well plate; rapidly growing cells...making sure we always maintain a backup replicates of each clone in case of contamination. We do not normally...assay plate. We generally freeze at least 2 ampules each of 4 clones with different levels of expression ...

  13. Transferable Skills: Negotiation

    Type
    Blog Post
    ...vacation (New York City or Hawaii?). So how can you approach improving this transferable skill? Negotiation...one of us wins, the other loses. Although this approach to negotiation is the most straightforward, it... a loser when negotiating, what if we instead approached it as a creative problem-solving exercise? Asking... an important plans? What is most important to each person? Someone has to be the first author but ...that the brand new grad student keeps using the machine? Let’s walk through a more specific example.... on many specific details about the individuals, each example provides a scenario where both people get...to be thoughtful about what is most important to each person. It can especially be difficult to think ...

  14. Common Injection Routes in Mice

    Type
    Blog Post
    ... do not require any punctures. Intratracheal instillation Intratracheal instillation is used to deliver...invasive. Restraint: anesthesia Location: trachea Gauge (tracheal tubing): 23–20 Volume: < 0.07 ml Skill ...Spinal cord 30 up to 0.01 mL Expert Intratracheal Anesthesia Trachea 20–23 (tubing) < 0.07 mL Advanced ...Intranasal instillation is very similar to its intratracheal counterpart, as both procedures target the lungs...experimental therapies or pain relief, straight to the stomach. This technique involves administering medication...

  15. Antibodies 101: Flow Compensation

    Type
    Blog Post
    ... do I fix it? When plotting two channels against each other, you should be able to draw perpendicular ...APC control, with APC and APC-CY7 plotted against each other. Since it is a single-color control, you know... Inputting compensation values differs between machines or the program you’re using, but usually you can...compensation. APC and APC-Cy7 are both bleeding into each other, as is evident by the two diagonal populations...populations are not perfectly straight, but the center of each population is in a line with the other populations... APC and APC-Cy7, with compensation panels below each plot.  The general rules of compensation apply... and APC should not require compensation between each other. We can therefore conclude that this population...

  16. Better Dyeing Through Chemistry & Small Molecule Fluorophores

    Type
    Blog Post
    ... “the photon budget”: each sample has a limited number of fluorophores and each fluorophore can emit a... a limited number of photons before bleaching. The amount of information one can extract from a biological...irreversibly with a small substrate ligand motif attached to a fluorophore (Figure 1b). Engineered ligases...phosphopantetheinyltransferase) – These enzymes catalyze the covalent attachment of a fluorophore ligand to a peptide tag (Figure...bioorthogonal chemical reactions (i.e., click chemistry) to attach a fluorophore at the specific site of incorporation... the speed and selectivity of the fluorophore attachment, the brightness of the resulting conjugate, and...budget gets a bit thin, small-molecule labeling approaches can provide a fresh infusion of photons for your...

  17. Microbes: Look, Listen, and Tell

    Type
    Blog Post
    ...acoustic reporter genes’ on our blog. Creative approaches to presenting the microbial world After learning...the lab, I was excited to learn about creative approaches to visualize bacteria in an educational setting...setting. The Symposium on ‘Creative Approaches to Presenting the Microbial World’ showcased complex connections...connections and ideas in various visualized approaches from hand-drawn comics (Dr. Sanja Saftic) to educational...makes scientific topics both interesting and approachable!   The grand finale of visual impressions was... creative ways to do science communication and outreach - if you'd like to learn more about Science communication... Susanna Bachle...

  18. Healthcare Consulting: A Door to the Business of Life Sciences

    Type
    Blog Post
    ... thanks for our guest blogger, Gairik Sachdeva! Gairik Sachdeva is a Ph.D. in Bioengineering and works...This post was contributed by guest blogger Gairik Sachdeva. Healthcare consulting is a fast-paced field, ...challenges, quickly breaking them up into smaller achievable tasks, and then executing on them in a timely...creative solutions to emerge. They enjoy the fact that each new challenge will require knowledge of new topics...situations, and personalities. Teams striving to achieve a lot of great output in a short amount of time... and to help manage that, most consulting firms coach and counsel employees on the different personality...core of the suite of benefits a given therapeutic approach brings to patients, doctors, and insurance providers...

  19. Analyzing CRISPR Editing Results with ICE from Synthego

    Type
    Blog Post
    ...then characterizes the sequence and abundance of each particular indel.   For a complete guide to ICE ... to gather details on the issues associated with each sample. Successfully analyzed samples will display...Sample - The unique label name that you provided for each sample. Guide Target - This is the user-defined ... name. You can perform more in-depth analyses on each sample by clicking on the sample name or on its ...analysis will open up a new window with three tabs.  Each of the three tabs - Contributions, Indel Distribution...entire edited population of genomes. Hovering over each bar of the indel plot shows the size of the insertion...base calls from the .ab1 file are also shown above each trace. The horizontal black underlined region represents...

  20. Advancing Biology with Zebrafish: Genetic Tools for Developmental Studies and More

    Type
    Blog Post
    ...investigations. Genetic resources for zebrafish A common approach to unravel the gene networks that drive biology... regions is also possible using an alternative approach that exploits the bacteriophage integrase phiC31...useful, I’ll outline two recent lineage tracing approaches that led to important insights in developmental...embryogenesis live. The origins, divisions, and movements of each of the thousands of cells in the developing zebrafish...tracked using microscopy and advanced computational approaches (Keller et al., 2008). This microscopy-based ... been mapped using single cell RNA sequencing approaches (Harland, 2018). Several different methods have...to manipulation of brain activity: optogenetic approaches can be used to determine the consequences of ...
Showing: 181 - 200 of 948 results