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Enter a sequence to perform a BLAST-based search of Addgene plasmid sequence databases.

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We narrowed to 1,019 results for: CAL;

Showing: 861 - 880 of 1019 results

  1. Plasmid Cloning by Restriction Enzyme Digest (with Protocols)

    Type
    Protocol
    ...treatment of your recipient plasmid as well as a specifically designed test digest later to verify that the...plasmid and 1μg of recipient plasmid. It is also critical that as much of the recipient plasmid as possible...step, depending on the phosphatase you choose. CIP (calf alkaline phosphatase) or SAP (shrimp alkaline phosphatase...base pairs for each varies, it is difficult to calculate this based on DNA concentration alone. One method...electro-competent cells instead of the more common chemically-competent cells. The number of bacterial colonies...will tell you your “background” level or more specifically it will tell you how many colonies you can expect...

  2. AAV ddPCR Titration

    Type
    Protocol
    ...Adeno-associated viral vectors (AAV). This protocol specifically uses primers and probes targeting the ITR elements... is genome copies and generally represents the physical number of viral particles. Users may need to run...biosafety regulations. Equipment Class II, Type A2 Biological Safety Cabinet, Labconco 302411100 Droplet Digital...cartridge holder with bleach and place in the Biological Safety Cabinet (BSC). Make sure that the BSC ...Pro-Tip Prepare the viral dilutions in a separate biological safety cabinet (dilution BSC) than that used ... a CSV file and use the following equation to calculate the titer for each sample: $$T = {R*C*D*1000\over...

  3. Weighing Reagents Protocol

    Type
    Protocol
    ...material, use an analytical scale. This will provide more accuracy as these types of scales display more ...are weighing by looking for a weight range on the scale. Make sure that the weight of the material you’re... balance. This will help you remove it from the scale later on as otherwise, the paper could stick to ...

  4. Centrifugation

    Type
    Protocol
    ...larger conical tubes. Depending on the centrifuge their speed ranges can vary, but are typically lower ... the two, use a conversion calculator or chart like the G Force calculator (Link opens in a new window...Tabletop Centrifuges sit on your benchtop and typically hold the common 1.5 mL microfuge tubes used in...

  5. Protocols for Molecular Biology, Plasmid Cloning, and Viral Preps

    Type
    Protocol
    ... Addgene Protocols On this page you'll find practical lab protocols that you can use for a wide range...Generating Stable Cell Lines with Lentivirus Genomically integrate your DNA sequence into a cell line ... SYBR Green Technology Use qPCR to measure the physical titer of your adeno-associated virus preparation...AAV Titration by ddPCR Use ddPCR to measure the physical titer of your adeno-associated virus preparation...Lentivirus Titration by ddPCR Use ddPCR to measure the physical titer of your lentivirus preparation Antibodies...

  6. Molecular Biology Protocol - Restriction Digest of Plasmid DNA

    Type
    Protocol
    ...depends on your application. A diagnostic digest typically involves ∼500 ng of DNA, while molecular cloning...largely determined by the volume of DNA to be cut. A typical restriction digestion reaction could look like ...reaction in 1 hour. Using this ratio, you can calculate the minimal amount of enzyme for your reaction...the vector if you are cloning in an insert. CIP (calf alkaline phosphatase) or SAP (shrimp alkaline phosphatase...enzymes cut sequences which are similar, but not identical, to their recognition sites. This is due to "...

  7. Transfection for Recombinant Antibodies

    Type
    Protocol
    ...significantly and should be empirically determined for your sample. Typical ratios may range from 1:1 to...pipettes Benchtop centrifuge compatible with 50 mL conical tubes Automated cell counter 37 °C, 5% CO 2 incubator...antibody plasmid DNA 500 mL vented flask 50 mL conical tubes Microcentrifuge tubes Cell counting chamber.... Transfer the HEK293 cells and media to 50 mL conical tubes. Centrifuge for 15 min at 3100 x g to pellet...

  8. What is Polymerase Chain Reaction (PCR)

    Type
    Protocol
    ...polymerase that can withstand radical temperature changes during a typical PCR. The DNA polymerase has ...template), a researcher can create thousands of identical copies using a simple set of reagents and a basic...GC-rich templates. What does each ingredient specifically do? Template DNA: Contains the portion of DNA...Buffer with MgCl 2 : Provides an optimal and stable chemical environment for the DNA polymerase to work adequately...

  9. Virus Protocol - Generating Stable Cell Lines

    Type
    Protocol
    ...stable cell lines Equipment Class II, Type A2 Biological Safety Cabinet 0.5–10 µL single channel pipette...mL), Millipore TR-1003-G 1X PBS pH 7.4 without calcium or magnesium, Corning 21-040-CV (cations can affect...tissue culture treated dish, Corning 3516 15 mL conical tubes, VWR 21008-216 Lentivirus preparation Appropriate...You Start The health of the target cell line is critical for obtaining accurate results. Check the cells...and/or harvest to test for protein expression. Typically, cells transduced with lower dilutions of the ...

  10. Gibson Assembly Protocol

    Type
    Protocol
    ...should have identical sequences on the ends (sequences A and B in the figures). These identical sequences...PCR with primers that contain a 5′ end that is identical to an adjacent segment and a 3′ end that anneals..., Hutchison CA, Venter JC, Merryman C. (2010). Chemical synthesis of the mouse mitochondrial genome. Nature...

  11. Protocol - Bacterial Transformation

    Type
    Protocol
    ...make bacteria more susceptible to either chemical or electrical based transformation, generating what are...antibiotic containing agar plate. This step is not critical for Ampicillin resistance but is much more important...large plasmids (>10 kb) or BACs, what can I do? Chemically competent cells are fast and easy to use, but...

  12. Protocol - How to Run an Agarose Gel

    Type
    Protocol
    ...visualization and purification. Electrophoresis uses an electrical field to move the negatively charged DNA through...eye protection and gloves when working with this chemical. Note: If you add EtBr to your gel, you will also...approximately 75-80% of the way down the gel. A typical run time is about 1-1.5 hours, depending on the...polyacrylamide gel electrophoresis (PAGE), which is typically used to separate 5 - 500 bp fragments. How do ...

  13. Protocol - How to Streak a Plate

    Type
    Protocol
    ...medium. This dot is composed of millions of genetically identical bacteria that arose from a single bacterium...draw in discontinuous lines. Start by streaking a vertical line of bacteria along one edge of the plate. ...

  14. Ligation Independent Cloning

    Type
    Protocol
    ...polymerase can create overhangs of varying length (typically 10-12 bp) based on a specific sequence. The annealed... the replication cycle. Empty vectors for LIC typically employ type II restriction enzymes (e.g. BsaI)...back the guanosine residue and become stalled. A typical T4 Pol reaction is shown. Mix the components (...

  15. Using a Light Microscope Protocol

    Type
    Protocol
    ...lab! Introduction Microscopes are emblematic of biological research and are found in many different types...suggests, light microscopes take advantage of the physical properties of light to detect small objects. Two...the light follows from the source to your eyes is called the light path . Light travels from the light source...

  19. Optogenetics Guide

    Type
    Guide
    ...and optical switches Sensors are genetically-encoded reporters of molecular signals; e.g., calcium indicators...Deisseroth K. 2005. Millisecond-timescale, genetically targeted optical control of neural activity. Nat... utilized to control the localization of a protein of interest. A genetically-encoded nuclear Cry2 fusion...MJ. 2006. Next-generation optical technologies for illuminating genetically targeted brain circuits. J...17375185 Kim B, Lin MZ. 2013. Optobiology: optical control of biological processes via protein engineering. Biochem... including explanations of microbial opsin and optical switch terminology. Science...Optogenetics Guide Introduction Microbial Opsins Optical Switches References Introduction The field of optogenetics...

  20. Antibody Guide

    Type
    Guide
    ...antibody’s constant region - specifically, it is determined by a region called the “heavy chain constant ...reagents are molecular tools used in the lab to specifically and tightly bind to other proteins. Antibodies...interest. The protein that an antibody recognizes is called an antigen, while the specific binding site is ...referred to as an epitope. Production Antibodies are typically produced in one of three different methods: in...independently manipulated. Antibodies produced this way are called recombinant antibodies. Recombinant antibodies ...are responsible for binding the antigen, and are called the antigen binding fragments (Fab). The tips of...the Y structure and the lower part of the arms is called the constant region, and is shown in dark blue ...
Showing: 861 - 880 of 1019 results