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We narrowed to 903 results for: Nes

Showing: 141 - 160 of 903 results
  1. Pipetting Protocol

    Type
    Protocol
    ...hundreds of µL, the second is tens, and the third is ones. Therefore 100 µL would read as 100 from top to ...P20: The first digit is tens of µL, the second is ones, and the third (red) digit is tenths. Therefore,...2.2 µL would read as 022. P2: The first digit is ones of µL, the second red digit is tenths, and the third...at the bottom. For the P1000, this represents the ones. For the P200, this represents the first decimal...
  2. Protocol - How to Streak a Plate

    Type
    Protocol
    ...Then streak horizontal lines in another section of the plate, and then diagonal lines in another section ...very popular technique is to draw in discontinuous lines. Start by streaking a vertical line of bacteria ...
  3. Protocol - How to Purify DNA from an Agarose Gel

    Type
    Protocol
    ...DNA in other lanes contaminating your sample. To accomplish this, it is best to skip lanes between samples...couple simple ways to increase the resolution (crispness) of your DNA bands include: a) running the gel...
  4. Protocol - How to Perform a Diagnostic Digest

    Type
    Protocol
    ...restriction enzyme -based cloning to test individual clones before use of more expensive forms of plasmid verification... want to know how to differentiate between two clones that differ only in the orientation of the insert...Simply run the digest products on a gel and those clones with the appropriately sized products should have...
  5. Video Library

    Type
    Protocol
    ...Addgene's Business Development Manager, discusses his transition from the Scientist Team to Business Team,...CRISPR Generate genomic deletions in mammalian cells lines using CRISPR/Cas9 Genomic Deletions with CRISPR ...
  6. Protocols for Molecular Biology, Plasmid Cloning, and Viral Preps

    Type
    Protocol
    ...the RNAi consortium for targeting human and mouse genes. Virus Protocols for titering and testing your virus...antibiotic resistant colonies Generating Stable Cell Lines with Lentivirus Genomically integrate your DNA sequence...Population by Limiting Dilution Generate monoclonal cell lines from a polyclonal pool of stable cells AAV Production...
  7. Fluorescence Titering Assay

    Type
    Protocol
    ... but can be adapted to a variety of target cell lines and selection markers. Note, because this protocol...Millipore TR-1003-G 1X PBS pH 7.4 without calcium or magnesium, Corning 21-040-CV (cations can affect the attachment...transduction efficiency. Titer will vary between cell lines. It is not recommended to subject lentiviral supernatants...
  8. Lentivirus Production

    Type
    Protocol
    ... can be modified for alternative packaging cell lines or transfection reagents. Once produced, lentivirus...Heat-inactivated FBS 1x PBS pH 7.4 without calcium or magnesium, Corning 21-040-CV (cations can affect the attachment... cells in a T75 flask in 15 mL DMEM Complete. Wednesday: Plate 1×10 6 cells in a T75 flask in 15 mL DMEM...
  9. Protocol - How to Run an Agarose Gel

    Type
    Protocol
    ...migrate through the gel more quickly than longer ones. Thus, you can determine the approximate length ...you can infer the size of the DNA in your sample lanes. For more details on doing diagnostic digests and...A few simple ways to increase the resolution (crispness) of your DNA bands include: a) running the gel...
  10. Coomassie Purity Stain of Recombinant Antibodies

    Type
    Protocol
    ... to the appropriate well. Pro-Tip Leaving clear lanes between samples will make quantifying the gel easier...Gels . Select Select Next Lane . Repeat for all lanes. Plot the area under the curves for the protein ...Figure 3 Select Analyze . Select Gels . Select Plot Lanes . One graph per lane will appear with peaks representing...
  11. AAV Production in HEK293 Cells

    Type
    Protocol
    ...Reagents Adherent HEK293T cells (ideally AAV-293T clones) Pro-Tip While adherent, these cells are very loosely...Fisher, 12605010) 1X PBS pH 7.4 without calcium or magnesium, Corning 21-040-CV (cations can affect the attachment... + 60 mL of 5 M Sodium Chloride + 4 mL of 1 M Magnesium Chloride Close the bottle and mix by inverting...
  12. CRISPR Library Amplification

    Type
    Protocol
    ...short region of DNA: guide RNAs targeting different genes in an organism's genome, for example. Amplification...altering the composition of the library. Bottlenecks, fitness differences, and plasmid recombination can all ...being lost from the pool and/or increasing the skewness of the pool. 8 electroporation cuvettes (BioRad...
  13. General Transfection

    Type
    Protocol
    ...This approach can be adapted for different cell lines and different transfection reagents. Workflow Timeline...cells in a 75 cm 2 flask in a volume of 15 mL. Wednesday: Plate 1x10 6 cells in a T75 flask in a volume...
  14. Immunocytochemistry

    Type
    Protocol
    ... fluorescent secondary antibody. This protocol outlines the steps for fixing and labeling HeLa cells for...and follow your institution's laboratory safety guidelines for disposing of waste in the appropriate container...
  15. Kit Free RNA Extraction

    Type
    Protocol
    ...dissociation of the nucleoprotein complexes. The effectiveness of your RNA isolation will depend on how effective...
  16. A Practical Approach to Choosing the B(right)est Fluorescent Protein

    Type
    Blog Post
    ...are (i) brightness, (ii) photostability and (iii) aggregation tendency. Brightness Brightness refers ...molecule. A higher brightness increases the detected signal and therefore high brightness is an important...how the brightness is best defined or determined. Here, we will explain how practical brightness is determined...determined and why practical brightness is a better criterion than theoretical brightness for selecting the brightest...protein. Theoretical brightness Two key properties of fluorophores that determine brightness are the extent ...number, the higher the theoretical brightness. The theoretical brightness often appears in tables with fluorescent...and the sample. Practical brightness A protein with a high intrinsic brightness that does not fold well ...
  17. Cloning Mammalian Cells with the Agarose Method

    Type
    Blog Post
    ... the Lindberg Lab we often make cell lines that overexpress genes of interest; more recently we have also...CRISPR vectors to generate cell lines with knockouts of specific genes. Many years ago, people in the ...give rise to single clones, and thus many plates must be examined for single clones and then handled. Additionally... Additionally, many cell lines, especially the endocrine cell lines we most commonly work with, require...cell clones with sterile truncated pipet tips. Indeed, an experienced operator can pick 48 clones from...individual cell clones with different expression characteristics from a population of transfected clones. The technique...number of single clones obtained (on occasion we have prepared two 48-well plates of clones) which enables...
  18. 27 Hot Plasmids from 2016

    Type
    Blog Post
    ...targeting their genes of interest to generate mAID fusions to endogenous genes. Cell lines with the appropriate...targeting a total of 20,121 genes within the human genome (the identity of the genes and target sequences can...continuous expansion of compatible backbones, promoters, and genes available to the community. If you ...identify both core and context-dependent human fitness genes, the laboratory of Jason Moffat at the University...cell models and identified ~4-5-fold more fitness-related genes than have been found in previous screens...uncharacterized genes that are conserved among apicomplexans and important for T. gondii fitness. Sixteen of...enable researchers to screen the entire genome for genes that regulate a wide variety of phenotypes, including...
  19. Custom CRISPR Screens & the Green Listed Software

    Type
    Blog Post
    ..., which genes are good candidates for being involved in the studied phenotype. For robustness of a screen...presenter then showed which genes they had generated knockout mice for (all of the genes involved as I recall...differentially expressed genes that are known drug targets, or all differentially expressed genes that code for ... if genes that you asked for were not found in the reference library. It’s not uncommon that genes have...5 guide RNAs (gRNAs) targeting each of the 1000 genes you’d like to investigate in your next CRISPR screen...CRISPR screens targeting a (long or short) list of genes provided by the user. This approach is useful for...investigator generates a cell population where all genes are knocked out at the population level, but each...
Showing: 141 - 160 of 903 results