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We narrowed to 950 results for: cha

Showing: 141 - 160 of 950 results

  1. Lentivirus Production

    Type
    Protocol
    ...0.5–10 µL single channel pipette 2–20 µL single channel pipette 20–200 µL single channel pipette 200–1000...1000 µL single channel pipette Ice bucket CO 2 incubator Pipet controller Hazardous waste container pH ... one suitable with your protocols. FBS can be purchased already heat inactivated or it can be inactivated...panels: bright field images; right panels: GFP channel images. Last reviewed on: August 2, 2023...

  2. Fluorescence Titering Assay

    Type
    Protocol
    ...0.5–10 µL single channel pipette 2–20 µL single channel pipette 20–200 µL single channel pipette 200–1000...1000 µL single channel pipette Ice bucket CO 2 incubator Pipet controller Hazardous waste container Inverted... one suitable with your protocols. FBS can be purchased already heat-inactivated, or it can be inactivated...

  3. General Transfection

    Type
    Protocol
    ...0.5–10 µL single channel pipette 2–20 µL single channel pipette 20–200 µL single channel pipette 200–1000...1000 µL single channel pipette Ice bucket CO 2 incubator Pipet controller Hazardous waste container Reagents... one suitable with your protocols. FBS can be purchased already head inactivated or it can be inactivated...

  4. Optogenetics Guide

    Type
    Guide
    ...of CrChR2 590 Channelrhodopsins: cation channels from other species VChR1 Channelrhodopsin from Volvox ... mutations 540 Channelrhodopsins: chloride channels iChloC Chloride-conducting channel, CrChR2 with mutations...optogenetics plasmids for your experiments. Channelrhodopsins Channelrhodopsins are a foundational optogenetics tool... direct stimulation of ion channels. Naturally occurring channelrhodopsins were discovered in the green...chloride channels, these variants result in the hyperpolarization of neurons. Examples of anion channel variants...Jaws Browse Halorhodopsin plasmids . Archaerhodopsins Archaerhodopsin-3 (Arch) from Halorubrum sodomense...Description Peak Response Spectra (nm) Channelrhodopsins: cation channels from Chlamydomonas reinhardtii ChR2...

  5. Isolating a Monoclonal Cell Population by Limiting Dilution

    Type
    Protocol
    ...0.5–10 µL single channel pipette 2–20 µL single channel pipette 20–200 µL single channel pipette 200–1000...1000 µL single channel pipette CO 2 incubator Pipet controller Hazardous waste container Microscope Hemocytometer... In the absence of selective medium, there is a chance that the integrated transgene could be lost through...

  6. CRISPR Guide

    Type
    Guide
    .... You can therefore change the genomic target of the Cas enzyme by simply changing the target sequence...surface-exposed positively-charged grooves on Cas9. gRNA binding induces a conformational change in Cas9, shifting...Fanton, A., Tycko, J., Hinks, M., Chandrasekaran, S. S., Perry, N. T., Schaepe, J., Du, P. P., Lotfy, P., Bassik...approach. Figure 1: Overview of the basic CRISPR mechanism Engineered CRISPR systems contain two components... In this section, we will introduce the basic mechanisms of CRISPR and some of the ways the CRISPR components...efficiency, or flexibility of the Cas enzyme. Basic Mechanisms of CRISPR The requirements for a CRISPR knockout...target DNA. Figure 2: Overview of the NHEJ repair mechanism Cas9 will only cleave a given locus if the gRNA...

  7. Immunocytochemistry

    Type
    Protocol
    ...0.5–10 µL single channel pipette 2–20 µL single channel pipette 20–200 µL single channel pipette 200–1000...1000 µL single channel pipette Reagents 1X PBS Microcentrifuge tubes Sterile Poly-D-lysine coated coverslips...

  8. AAV Purification by Iodixanol Gradient Ultracentrifugation

    Type
    Protocol
    ... Nonetheless, we recommend performing a buffer exchange before using the purified AAV in vivo . The iodixanol... Workflow Timeline Day 1: Purify Day 2: Buffer exchange and concentration Note: Both steps could be completed...Pool clean fractions. Concentration and buffer exchange: Stocks of Pluronic F68 Pluronic F68, 10% solution...Choi, and R. Jude Samulski. "Production and characterization of adeno-associated viral vectors," (Link ...

  9. Antibody Guide

    Type
    Guide
    ...made up of four protein chains: two large heavy chains and two small light chains. These four domains come... producing antibodies, can change the constant region, and therefore change the isotype of antibodies ...: scFv - Single-chain fragment variables (scFvs) contain only the variable heavy chain and variable light... it is determined by a region called the “heavy chain constant region” or Fc. Different isotypes play ...light chain regions of an antibody. They are small enough to be produced in bacteria (~25 kD), but tend... derived from a camelid antibody called a heavy chain antibody (HCAb). Their small size allows them to... conjugation reaction. Many antibodies can be purchased already conjugated, which can save significant...

  10. Delivery Methods for Generating iPSCs

    Type
    Blog Post
    ...Young-Gie Chung, Mi-Yoon Chang, Baek-Soo Han, Sanghyeok Ko, Eungi Yang, Kwang Yul Cha, Robert Lanza, and Kwang-Soo...Plasmids for Stem Cell Research Delivery Method Mechanism Efficiency Safety Pros Cons MMLV-derived Retrovirus...element that transposes via a “cut and paste” mechanism between a vector and chromosomal DNA. PB has a...be removed by re-expressing the transposase. A challenge of using PB to generate iPSCs is that genomic ...the cytoplasm independent of the cell cycle. A challenge of using Sendai is that since it’s replication...clones tested had plasmid integration. Another challenge is that multiple transfection makes it difficult...plasmids due to lower activation of DNA-silencing mechanisms. While minicircles are typically smaller than...

  11. Chemogenetics Guide

    Type
    Guide
    ... specific cellular pathways, receptors, or ion channels targeted to specific cell populations (most often...control of neurons through manipulation of ion channels arePharmacologically Selective Actuator Modules...ion pore domain (IPD) to form ligand-gated ion channels (LGICs) that can either excite or inhibit neuronal...activity. Figure 3: PSAM4-based ligand-gated ion channels, their effect, and outcome in neurons. Table 2...opens in a new window) Coward P, Wada HG, Falk MS, Chan SD, Meng F, et al. (1998). Controlling signaling..., Harvey BK, Dannals RF, Pomper MG, Bonci A, Michaelides M (2017). Chemogenetics revealed: DREADD occupancy...Ikefuama EC, Slaviero AN, Silvagnoli AD, Crespo EL, Schalau R, Gott M, Tree MO, Dunbar GL, Rossignol J, Hochgeschwender...

  12. Lentiviral Vector Guide

    Type
    Guide
    ... rev Envelope Plasmid Interchangeable; usually encodes for VSV-G Interchangeable; usually encodes for ...immunodeficiency virus (FIV). Retroviruses are partially characterized by their ability to integrate their genomes ...them into the supernatant. After an initial media change, the supernatant containing the viral vectors is...lentiviral vector will infect. This can be altered by changing the envelope gene, a process called pseudotyping...Lentiviral Vector Uses and Overview Overcoming the Challenges of Lentiviral Production Your Lentiviral Plasmid...doi.org/10.1126/science.272.5259.263 PMID: 8602510 Schambach, A., Zychlinski, D., Ehrnstroem, B., & Baum, C...10.1261/rna.2192803 PMID: 12649500 Timms, R. T., Tchasovnikarova, I. A., & Lehner, P. J. (2018). Differential...

  13. Adeno-associated virus (AAV) Guide

    Type
    Guide
    ...construction, as it was the first to be well-characterized and extensively studied, and has a broad tissue...of these hybrid viral vectors have been well characterized and are often preferred over standard rAAV vectors...response to light. The short coding sequences of channelrhodopsins, halorhodopsins, and other optogenetic genes...) and a gRNA together into an rAAV capsid is challenging due to its limited packaging capacity, especially...later reconstructed within the cell via different mechanisms. One of these methods uses splice site donor ...window) PMID: 15294177 (Link opens in a new window) Chan, K. Y., Jang, M. J., Yoo, B. B., Greenbaum, A., ...12663794 (Link opens in a new window) Kwon, I., & Schaffer, D. V. (2008). Designer gene delivery vectors:...

  14. Gamma-Retroviral Vector Guide

    Type
    Guide
    ...leukemia virus (FeLV). Retroviruses are partially characterized by their ability to integrate their genomes ...gamma-retroviral production, and scientists have made changes to these components for increased safety when producing...into distinct plasmids for safety to reduce the chance of creating replication-competent retroviruses.... modify tropism, as the env gene can easily be changed. The HEK293T cells will produce the virus and release...it into the supernatant. After an initial media change, the supernatant containing the viral particles...gamma-retroviral vector will infect. This can be altered by changing the envelope gene, a process called pseudotyping... 24464997 Maetzig, T., Galla, M., Baum, C., & Schambach, A. (2011). Gammaretroviral vectors: Biology, ...

  15. Molecular Biology Protocol - Restriction Digest of Plasmid DNA

    Type
    Protocol
    ...restriction digest results. Equipment Electrophoresis chamber Pipetman Reagents Liquid DNA aliquot of your plasmid...enzymes. Most companies will have a compatibility chart, such as the (Link opens in a new window) double...in a new window) NEB's compatible cohesive ends chart . If you cannot find compatible sticky ends, you...

  16. Transfection for Recombinant Antibodies

    Type
    Protocol
    ... blue mix into a cell counting chamber. Load the cell counting chamber on an automated cell counter and...conical tubes Microcentrifuge tubes Cell counting chamber slide 30 mL luer-lock syringe, BD BD302832 0.2 ...

  17. Protocol - How to Run an Agarose Gel

    Type
    Protocol
    ...uses an electrical field to move the negatively charged DNA through an agarose gel matrix toward a positive...some to the buffer as well. EtBr is positively charged and will run the opposite direction from the DNA...negative, red is positive. The DNA is negatively charged and will run towards the positive electrode. Always...

  18. Guide to Using Pooled Libraries

    Type
    Guide
    ...numbers of cells minimizes the effects of random chance that could lead to false positive or negative results...being studied. Examples of published selection mechanisms for lentiviral CRISPR libraries include: resistance...that disappear with addition of the selection mechanism Positive Screen In a positive screen, the goal...those cells that do not survive the selection mechanism. You’ll infect two sets of cells and subject one...consult these links: NGS Company Resources Science Exchange Scientist (formally Assay Depot) Resources Find...

  19. CRISPR Library Amplification

    Type
    Protocol
    ...Quality Control (QC) Pooled libraries can be challenging and expensive to ensure adequate quality, but...can be used but this dramatically increases the chances of individual plasmids being lost from the pool...taken to perform adequate NGS based QC to ensure no change in representation compared to the pre-amplified...

  20. Adenovirus Guide

    Type
    Guide
    ...provide the deleted adenoviral genes. The biggest challenge of the HCAdV vector production is to ensure no...Ebola virus, and Zika virus. Chimpanzee adenoviral (ChAdV) vectors have also been developed as vaccine candidates...window) PMID: 21453277 (Link opens in a new window) Bulcha, J. T., Wang, Y., Ma, H., Tai, P. W. L., & Gao,...., Pierson, T. C., & Weaver, E. A. (2020). Characterization of a Species E Adenovirus Vector as a Zika... protect nonhuman primates against ebolavirus challenge . Journal of Virology, 85 (9), 4222–4233. https...
Showing: 141 - 160 of 950 results