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We narrowed to 616 results for: TIL

Showing: 41 - 60 of 616 results

  1. Using a Light Microscope Protocol

    Type
    Protocol
    .... You can take entire courses on microscopy and still have more to learn, so, for this protocol, we'll...adjustment knob (the larger of the two focus knobs) until you can see the edges of the aperture are in focus...field of view and then slowly open the diaphragm until the edges are just out of view. Pro-Tip Looking ...arm to move the stage (and ultimately your slide) until your field of view is centered on a region of interest...

  2. Protocol - How to Run an Agarose Gel

    Type
    Protocol
    ...buffers and do not use water). Microwave for 1-3 min until the agarose is completely dissolved (but do not ... OR let sit at room temperature for 20-30 mins, until it has completely solidified. Pro-Tip If you are...electrophoresis unit). Fill gel box with 1xTAE (or TBE) until the gel is covered. Pro-Tip Remember, if you added...additional wells of the gel. Run the gel at 80-150 V until the dye line is approximately 75-80% of the way ...

  3. Kit Free RNA Extraction

    Type
    Protocol
    ... Work in a well-ventilated space and under a fume hood when working with the volatile reagents in the ...entire tube is dried but while the white pellet is still visible. Resuspend RNA pellet in RNase-free water...

  4. Protocol - pLKO.1 – TRC Cloning Vector

    Type
    Protocol
    ...digested DNA on 0.8% low melting point agarose gel until you can distinctly see 2 bands, one 7kb and one ... replace with fresh media. Do not add puromycin until at least 24 hours after infection to allow for sufficient...guidelines for the number of days you should wait until harvesting your cells. However, you should optimize...solution by dissolving 40g of LB powder in 1L of distilled water. Autoclave and cool to 55°C. Add 1mL of ...of puromycin (Sigma-Aldrich catalog #P8833) in distilled water. Sterilize by passing through a 0.22 μm ...

  5. Water Bath Protocol

    Type
    Protocol
    ...Equipment Water Bath 10% bleach or 70% ethanol Distilled water Disinfectant Thermometer Water bath weights...the edge of the slide. Fill the water bath with distilled water, not tap water, to prevent salts from accumulating...water in it from previous uses. However, you will still need to regularly clean out the water bath even ...

  6. Western Blot

    Type
    Protocol
    .... Use the lysate immediately or store at -80 °C until ready to use. Section 2: Determine the total protein...power supply. Run the gel at 100 V for 10–15 min or until the samples have moved out of the wells and into...the voltage to 150 V and continue running the gel until you have obtained the desired separation. Gently...

  7. AAV ddPCR Titration

    Type
    Protocol
    ...on the chilled 96-well freezer block in the BSC until ready to use. Prepare the Master Mix Place a ddPCR...pipette tips directly in the center of the well and tilt to a 45° angle. Count to 20 while slowly and gently...Lift the tips ~1 mm. Touch the side of the well and tilt the pipette tips at a 45° angle. Count to 20 while...

  8. Lentivirus ddPCR Titration

    Type
    Protocol
    ... used for ddPCR immediately or stored at -20 °C until ready to use. Preparing for ddPCR Thaw samples, ...pipette tips directly in the center of the well and tilt to a 45° angle. Count to 20 while slowly and gently...Lift the tips ~1mm. Touch the side of the well and tilt the pipette tips at a 45° angle. Count to 20 while...

  9. 15 Hot Plasmids from 2017

    Type
    Blog Post
    ...recently deposited 2 Bacillus subtilis libraries with Addgene – the B. subtilis Single Gene Deletion Library—Kanamycin...target specific neuronal subtypes was not available until now. In order to target specific neuronal subtypes...to light. Robert Campbell’s lab illustrates the utility of PhoCl in a variety of settings. For instance...the podcast Segement on the pPSU Plasmids B. subtilis knockout and knockdown libraries The Gross Lab... Essential gene Knockdown library. Although B. subtilis has a fully sequenced genome, many applications... to advance our collective understanding of B. subtilis gene functions and interactions. The single-gene...single-gene deletion library is derived from the B. subtilis 168 strain and contains knockouts for all 3984 genes...

  10. Getting to Know Expansion Microscopy

    Type
    Blog Post
    ...break through the resolution barrier while still utilizing fluorescence microscopy, is another option....more accessible imaging equipment, ExM protocols utilize relatively inexpensive reagents and are readily...fluorescent proteins (Gambarotto, et al. 2019; Tillberg, et al. 2016). That being said, the anchoring,... between them to ensure the expanded images are still a good representation of reality. Third, expansion...secondaries, can improve signal strength (Sherry and Stiles, 2022). Fourth, signal continuity can appear reduced.../doi.org/10.1371/journal.pbio.3001020  Chen F, Tillberg PW, Boyden ES (2015) Expansion microscopy. Science...https://doi.org/10.1098/rsta.2021.0110  Sherry DM, Stiles MA (2022) Improved fluorescent signal in expansion...

  11. Protocol - Over-Agar Antibiotic Plating

    Type
    Protocol
    ... of the agar and gently spread over the surface until the liquid is mostly absorbed (there is a very small...onto the agar and gently spread over the surface until the liquid is mostly absorbed. The spreading of ...

  12. Plasmids 101: Yeast Vectors

    Type
    Blog Post
    ...within an E. coli cell, but vectors can be widely utilized across many different cell types and each one ...can post-translationally modify proteins yet they still share many of the same technical advantages that..., a well-defined genetic system, and a highly versatile DNA transformation system.Unlike most other microorganisms...of these vectors. S. pombe plasmids oftentimes utilize an ARS to aid in high transformation efficiency...markers for yeast Historically, scientists have utilized auxotrophic selection rather than antibiotic selection...auxotrophic strain/ selection element pairs must be utilized or a new combination needs to be created in advance...improvements in antibiotic selection have made utilizing the more traditional drug selection methods feasible...

  13. Ligation Independent Cloning

    Type
    Protocol
    ...will remove bases from the 3' end of the cut site until the first G is reached (shown in blue), at which...causing the enzyme to perform exonuclease activity until the first "G" in the sequence. Because the polymerase...

  14. Transferable Skills Guide: Time Management

    Type
    Blog Post
    ...have waited. Both tasks have some utility, but I lost all the utility (fun) of eating with friends by splitting... meeting up with friends, but didn’t realize it until it was too late. I didn’t spend enough time thinking... two main things: How useful the task is (its utility) How much time it will take you to complete the...should err on the side of things that have the most utility; i.e. those things that will be most useful for...option. Even if the full experiment fails, you’ll still have made small accomplishments each day that should...and think about what they mean in terms of the utility of your tasks. In my cell splitting anecdote, I...hood may have prevented me from splitting cells until late in the day anway. There are a few things I’...

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

    Type
    Blog Post
    ...FLEx vector design Recombinase-dependent vectors utilize pairs of specific sequences, lox (recognized by...expression, low levels of spurious expression were still observed in many systems. So what is the cause of...of DIO/FLEx vectors, the cause of these events, until recently, was not known. A recent paper from Ed ...post-translational modifications, and other factors that AAV utilizes to gain entry into the cell (Dudek, AM. 2018)....in order to minimize off-target expression while still achieving adequate expression in your cells of interest...recombination rate (0.1% and 0.13% respectively) still required a 200-300 fold dilution to minimize off-target...accuracy and reproducibility of the data and can instill greater confidence when interpreting the outcome...

  16. General Transfection

    Type
    Protocol
    ...water. While stirring, slowly add hydrochloric acid until the solution clears. Check the pH of the solution...dislodge the cells. Incubate the cells for 18 h, or until the following morning. The following morning, carefully...

  17. 5 Reasons to Use Reddit for Science Communication

    Type
    Blog Post
    ...joining the conversation on Reddit, but it wasn’t until one of our blog writers posted his article (Developing...00 am and Joanne didn’t have to start answering until 5 pm). The ability to see the questions first makes...that I didn’t have the direct link to send people until the day of the AMA. This made it a little difficult...alter my intro to reduce the number of potentially hostile posters (this advice included leaving out the term...career path development--but the science trainees still have to lead on their own development. Please see...early in the day. Although I didn’t start answering until 5pm, frankly, I was typing far before five. Fortunately...your own professional development etc. database. Still have questions? I am easy to reach and happy to ...

  18. The CRISPR Software Matchmaker: A New Tool for Choosing the Best CRISPR Software for Your Needs

    Type
    Blog Post
    ... engineering. When compared to that collective utility, it’s easy to fault the lack of features in a single... can be used to aid design. These databases are still young and will require a collaborative effort from...arbitrary PAM definitions. These newer tools may still prove somewhat useful for the latest NTT targeting...protein, Cpf1. Separately, on-target efficacy is still a concept that only a few tools are trying to solve... workbenches such as MacVector or Benchling. Utility functions: These functions help speed up the sgRNA...single-method design”. Trends: The most common utility functions are batch design or multiplex features...use. The R based package CRISPRseek offers great utility and coupled with Bioconductor should be the first...

  19. Lentivirus Production

    Type
    Protocol
    ...water. While stirring, slowly add hydrochloric acid until the solution clears. Check the pH of the solution...disturb the cells. Incubate the cells for 18 h, or until the following morning. The following morning, carefully...

  20. Pipetting Protocol

    Type
    Protocol
    ...large container holding a small volume of liquid, tilt the container holding the liquid so that it's easier...second stop to release the liquid. Pro-Tip Dispense until the first stop, wait, and then dispense to the second...
Showing: 41 - 60 of 616 results