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Showing: 991 - 1020 of 1027 results

  1. Coomassie Purity Stain of Recombinant Antibodies

    Type
    Protocol
    ... Recombinant Antibodies Recombinant Antibody Purification Protocol Introduction This protocol describes...mobility of the bands (sample AR0016 in Figure 1) indicates that the samples may not have been processed correctly...ImageJ. Select File . Select Open . Choose the location of the file to open. Change the image type to ...select the Chart Type drop down menu and select Scatter chart. Change the data range and select the cells...

  2. Protocol - How to Perform a Diagnostic Digest

    Type
    Protocol
    ... . The pattern of the fragments on the gel can indicate if the plasmid contains the expected size insert...before use of more expensive forms of plasmid verification, such as DNA sequencing . In the example above...side of the insert, you can get a very obvious verification of the orientation so long as the expected products...

  3. Lentivirus Production

    Type
    Protocol
    ...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...inhibit transfection; therefore, plasmid DNA purification should include an endotoxin removal step. For...

  4. Colony Formation Titering Assay

    Type
    Protocol
    ...without calcium or magnesium, Corning 21-040-CV (cations can affect the attachment of adherent cells) 0.1%...Figure 1: A549 cells were transduced with the indicated serial dilutions of the lentiviral vector pRosetta...Figure 2: A549 cells were transduced with the indicated serial dilutions of the lentiviral vector pHAGE...

  5. Pipetting Protocol

    Type
    Protocol
    ...dispensed. The boxes that the tips come in often indicate a volume range that the tip can hold. This should...no liquid should drip from the tip. This could indicate that the tip is not on the pipette properly. Place...pipette tip by holding the pipette tip over your dedicated waste container and pressing on the tip ejector...

  6. Virus Protocol - Generating Stable Cell Lines

    Type
    Protocol
    ...without calcium or magnesium, Corning 21-040-CV (cations can affect the attachment of adherent cells) 0.45...lower dilutions depending on your downstream applications. If you’ve titered your virus beforehand, you...that the transgene has integrated in different locations in the various cells in the culture. This is because...

  7. Pouring LB Agar Plates

    Type
    Protocol
    ...Tetracycline 10 mg/mL 10 µg/mL Notes: Unless otherwise indicated, the antibiotic powder can be dissolved in dH ...test results. Sample Data In all cases below (-) indicates that the tested strain is not supposed to be resistant... resistant to the antibiotic, (+) indicates that the tested strain is supposed to be resistant to the ...

  8. Protocol - How to Design Primers

    Type
    Protocol
    ...primers produce inaccurate, nonspecific DNA amplification product, and long primers result in a slower...which creates primer dimers and disrupts the amplification process. When designing, if unsure about what...

  10. Fluorescence Titering Assay

    Type
    Protocol
    ... these cells can still be used in downstream applications. Safety Warnings Lentivirus is generally considered...without calcium or magnesium, Corning 21-040-CV (cations can affect the attachment of adherent cells) 0.45...

  11. Lab Safety for Biosafety Levels One and Two

    Type
    Protocol
    ... room. Make sure that you know where these are located before you start. Use the eyewash station if unwanted...choose to reuse or repurpose this SOP in another location, please note that you do so at your own risk; ...

  12. Immunocytochemistry

    Type
    Protocol
    ...Guide Western Blot Protocol Recombinant Antibody Purification Protocol Introduction Immunocytochemistry is...as methanol or acetone may be better for some applications. Remove the paraformaldehyde and follow your...

  13. Protocol - How to Run an Agarose Gel

    Type
    Protocol
    ... length in base pairs) for visualization and purification. Electrophoresis uses an electrical field to...instructions on how to do this, visit the Gel Purification page. Tips and FAQ How do you get better resolution...

  14. Antibody Validation Using the Indirect ELISA Method

    Type
    Protocol
    ...Immunocytochemistry Protocol Recombinant Antibody Purification Protocol Antibody Guide Introduction This protocol...display a green product. A precipitated product indicates the presence of too much HRP and the need to optimize...

  15. Isolating a Monoclonal Cell Population by Limiting Dilution

    Type
    Protocol
    ...without calcium or magnesium, Corning 21-040-CV (cations can affect the attachment of adherent cells) Trypsin...orange/yellow if using a standard phenol red pH indicator) because the buildup of waste products could be...

  16. AAV ddPCR Titration

    Type
    Protocol
    ...post on ddPCR for AAV quantification. This protocol was modified from the publication Lock et al. (2014) ...

  18. Weighing Reagents Protocol

    Type
    Protocol
    ...in the photo above has a capacity of 200 g, as indicated by the number above the "Zero" button. Before ...

  19. Water Bath Protocol

    Type
    Protocol
    ...bring your materials to a particular temperature, catalyze chemical reactions such as restriction digests...

  20. Centrifugation

    Type
    Protocol
    ...centrifuge begins to wobble or shake, this could indicate an unbalanced or improperly loaded centrifuge....

  21. General Transfection

    Type
    Protocol
    ...inhibit transfection; therefore, plasmid DNA purification should include an endotoxin removal step. For...

  22. Gibson Assembly Protocol

    Type
    Protocol
    ...product, gel-purify DNA segments. Otherwise, PCR purification or even the raw PCR mix can work fine in an ...

  23. CRISPR Guide

    Type
    Guide
    ... organisms, but modifications to various Cas enzymes have extended CRISPR applications to increasingly...following the amplification protocol specified by the depositing lab. After amplification, you’ll need ...heritability. Not all modifications are stable across cell divisions, and some types of modifications are more frequently... subsequently isolated by affinity purification. After purification, molecules associated with the locus...conventional methods for genomic purification, CRISPR-based purification methods are more straightforward...and SHERLOCKv2) allows for greater amplification and quantification and can be used on lateral flow strips...table above, 3' and 5' indicate on which end of targeted sequence the PAM is located. The majority of the...

  24. Antibody Guide

    Type
    Guide
    ...Handling Visualization Signal Amplification Application Overview Quantification Methods Capture Methods Visualization...Antibody Applications Antibody-based applications can be generally classed into four categories: Quantification... Read on to learn more about the applications common to each category, including an outline of the basic...these applications, visit the Antibody section of the Addgene Protocols page. Antibody Applications - Quantification...concentrates in the sample. Controls for quantification methods Quantification methods should be run with: a positive...using sonication to break DNA up into fragments of 300-1000 bps in length. Note: This sonication process...production and storage techniques, and explain common applications. Science...

  25. Adeno-associated virus (AAV) Guide

    Type
    Guide
    ...potential formation of replication-competent viruses. While AAV themselves are replication incompetent, there...production of replication-competent viruses that would be able to infect cells and replicate autonomously... actively causing symptoms or replicating. Lytic cycle Viral replication process where the virus uses ...into cells. Replication-competent rAAV Mutated rAAV vector that has the ability to replicate and reproduce.... The Rep region encodes four non-structural replication proteins (Rep78, Rep68, Rep52, and Rep40), which...which are involved in genome replication and viral packaging. The Cap region encodes three structural ...virus. The ITR sequences serve as origins of replication and also act as signals for packaging. Figure...

  26. Molecular Biology Reference

    Type
    Guide
    ... Origin of Replication ( ori ) DNA sequence which directs initiation of plasmid replication (by bacteria... Educational Resources Molecular Biology Reference Molecular...plasmid selection DNA sequencing for plasmid verification Resources Genetic Code Webpage and Blog References...Techniques in chemistry enable isolation and purification of cellular components, such as DNA, but practically...double-stranded DNA that typically carry genes and can replicate independently from chromosomal DNA. Although they...the bacterial chromosomal DNA, plasmid DNA is replicated upon cell division, and each daughter cell receives...manipulating genes. For instance, most plasmids are replicated in E. coli and are relatively small (∼3000 - ...

  27. Adenovirus Guide

    Type
    Guide
    ... DNA replication and host cell manipulation. Late transcripts are expressed after DNA replication starts...production of replication-competent adenoviruses (RCAs) that would be able to infect cells and replicate autonomously..., which has limited their use in therapeutic applications. However, this has also made them the most widely...presence of adenoviral genes E1, E4, E2a and VA for replication. For more information about AAVs, read our AAV...whereas the E1 region is essential for viral replication and therefore needs to be supplied through other... other early gene regions, E2 and E4. These modifications aimed to lower immunogenicity issues observed...also resulted in a reduction of viral vector amplification and overall lower viral titers. Because of this...

  28. Lentiviral Vector Guide

    Type
    Guide
    ...recently involved in translocation events, and other “fragile” genomic locations, and that this preference...therapeutic applications, like chimeric antigen receptor T cell (CAR-T) therapy, the location of integration...and tropism. Replication-competent The ability of a virus (or viral vector) to replicate and reproduce...hosts in order to continue their lifecycle and replicate. This feature can be harnessed for stable delivery...of safety improvements and the potential for replication-competent lentivirus production — that is, creation...particles that could infect cells and further replicate on their own. While the viral components are still...Interchangeable; usually encodes for VSV-G Safety Safe; replication incompetent by using three separate plasmids ...

  29. Gamma-Retroviral Vector Guide

    Type
    Guide
    ...and tropism. Replication-competent The ability of a virus (or viral vector) to replicate and reproduce...hosts in order to continue their lifecycle and replicate. This feature can be harnessed for stable delivery... for safety to reduce the chance of creating replication-competent retroviruses. For a summary of all ... gamma-retroviral vectors integrate at random locations throughout the genome. Gamma-retroviral vectors...retroviral vectors are commonly used in gene therapy applications to more specifically direct delivery of gene...vectors are a popular choice in gene therapy applications. There are many approved gamma-retroviral gene...cell (CAR-T) therapy is a particularly popular application of gamma-retroviral vectors. Gamma-retroviral...

  30. Optogenetics Guide

    Type
    Guide
    ...Channelrhodopsins: cation channels from Chlamydomonas reinhardtii ChR2 Widely used light-gated cation channel from...light of the correct wavelength, the pore opens, cations flow into the cell (yellow dots), and the neuron...reporters of molecular signals; e.g., calcium indicators In this overview, we will focus on the common... electrochemical responses, such as nonspecific cation influx vs. proton efflux. Researchers have used...is excited by blue light and permits nonspecific cation influx into the cell when stimulated. Channelrhodopsin...optogenetic tool, is also a blue light activated cation channel. ChR2 is preferred over ChR1 because ChR2...been expanded by scientists through both the identification of novel ChRs from other algal species and ...
Showing: 991 - 1020 of 1027 results