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We narrowed to 950 results for: omp

Showing: 141 - 160 of 950 results

  1. Plasmid Modification by Annealed Oligo Cloning (with Protocols)

    Type
    Protocol
    ...CAATTG ). The bottom oligo will be the reverse compliment so that they can anneal. Top oligo: 5' - CATATG...bp We also need to include additional bases to complement the overhangs generated when digesting the vector...could leave off the 3’ G on each oligo (and the complementary C of the other oligo), but this would destroy...following oligos from your favorite oligo synthesis company: Top oligo: 5' - AATTCCATATGTTAATTAAGGCGCGCCCAATTGG...annealed oligos). Transform 2-3μL into your favorite competent bacteria and plate. Be sure to pick multiple colonies...

  2. Fluorescence Titering Assay

    Type
    Protocol
    ...biosafety level 2+. Please ensure that you are in compliance with your institution’s biosafety regulations...Lentivirus preparation Reagent Preparation DMEM Complete: 10% v/v FBS and 4 mM L-alanyl-L-glutamine (or...follows: Dilute 525,000 cells into 14 mL of DMEM complete. Mix well by pipetting or inverting. Aliquot 2... Prepare dilutions of the lentivirus into DMEM complete containing 10 μg/mL polybrene. Note, this protocol...Volume of Lentivirus Stock (μL) Volume of DMEM complete (μL) Volume of 10mg/mL polybrene (μL) 1:10 150...

  3. General Transfection

    Type
    Protocol
    ...DMEM complete containing 25 µM chloroquine diphosphate and incubate ~5 h For 10 mL of DMEM complete, add...Syringes for filtering Reagent Preparation DMEM Complete: 10% v/v FBS and 4 mM L-alanyl-L-glutamine (or...HEK293T cells at 3.8x10 6 cells per plate in DMEM complete in 10 cm tissue culture plates. Incubate the cells...the media. Replace the media with 15 mL of DMEM complete. Incubate the cells 24–48 h before checking for...

  4. Affinity Purification of Recombinant Antibodies with Protein A or Protein G

    Type
    Protocol
    ...Refrigerator Pipette controller Benchtop centrifuge compatible with 50 mL conical tubes NanoDrop spectrophotometer... 250 mL sterile bottle, VWR 430281 0.45 µm PES complete filtration unit, 500 mL, rapid-flow, VWR 73520...Place a mark on the side of the column where the compacted resin is slanted upward. Place column in centrifuge...centrifugation step Pro-Tip Resin will appear compacted after centrifugation. Discard the flow through.... Slowly apply the sample to the center of the compact resin bed. Pro-Tip For sample volumes <1.5 mL top...

  5. What is Polymerase Chain Reaction (PCR)

    Type
    Protocol
    ...denaturation, annealing, and synthesis. After the PCR is complete, the product can be verified based on size by ...Diluting Primers Most people order primers from a company, which synthesizes and ships them as a lyophilized...essential that the template DNA and primer are complementary for a PCR to work efficiently. Taq Buffer with...Forward and Reverse Primers: Hybridize and are complementary to the 3’ ends of the sense and anti-sense strands...

  6. Isolating a Monoclonal Cell Population by Limiting Dilution

    Type
    Protocol
    ...Corning 430167 (optional) Reagent Preparation DMEM Complete: 10% v/v FBS and 4 mM L-alanyl-L-glutamine (or...Each 10 cm dish should be seeded in 10 mL DMEM complete, which will generate enough conditioned medium...conditioned medium, prepare this cell solution in fresh complete medium. This 5 cells/mL solution will be used ...adding 10 µL homogenized cell solution to 990 µL complete medium. Instead of transferring 0.125 µL, now ...

  7. AAV ddPCR Titration

    Type
    Protocol
    ...layout is complete, select 'Run' to begin the droplet reading. When the droplet reading is complete, export... on the insert. Please ensure that you are in compliance with your institution’s biosafety regulations...Stabilization 98 10 2 1 Hold 4 ∞ 2 1 After PCR is complete, transfer the plate to the Droplet Reader. Open...

  8. Ligation Independent Cloning

    Type
    Protocol
    ...restriction digest protocol ). After the digest is complete, you will need to separate the linearized vector... A typical T4 Pol reaction is shown. Mix the components (add the T4 last) and incubate at room temperature...of T4 Pol in the following step. This may be accomplished by gel purification . Step 5: Create Insert ...

  9. Centrifugation

    Type
    Protocol
    ... and using a centrifuge to separate different components in a liquid sample....laboratory equipment used to separate different components in a liquid sample. It achieves this by using... you use, most will have essentially the same components. The photo to the left shows a front view of ...

  10. Gibson Assembly Protocol

    Type
    Protocol
    ...2009). Regardless of fragment length or end compatibility, multiple overlapping DNA fragments can be joined...overhangs by chewing back from the DNA 5’ end. Complementary DNA fragments can subsequently anneal to each... DNA Ligase - covalently joins the annealed complementary DNA fragments, removing any nicks and creating...

  11. Protocol - How to Create a Bacterial Glycerol Stock

    Type
    Protocol
    ...bacterial strain and you will not need to obtain more competent cells and retransform. Bacteria on an LB agar ...streaking onto LB agar will prevent it from thawing completely and will improve the shelf life. It is very important...

  13. Transfection for Recombinant Antibodies

    Type
    Protocol
    ...biosafety level 2. Please ensure that you are in compliance with your institution’s biosafety regulations...tips Pipettor and pipettes Benchtop centrifuge compatible with 50 mL conical tubes Automated cell counter...

  14. AAV Purification by Iodixanol Gradient Ultracentrifugation

    Type
    Protocol
    ...vivo . The iodixanol gradient in this protocol is composed of steps that separate out contaminants from an...exchange and concentration Note: Both steps could be completed in one (long) day. Equipment Ultracentrifuge T70i...

  15. Protocol - How to Run an Agarose Gel

    Type
    Protocol
    ...). Microwave for 1-3 min until the agarose is completely dissolved (but do not overboil the solution, ...room temperature for 20-30 mins, until it has completely solidified. Pro-Tip If you are in a hurry, the...

  16. Handling Plasmids from Addgene - Purifying Plasmid DNA

    Type
    Protocol
    ...bacterial cultures , please see those pages. Several companies, such as (Link opens in a new window) NEB , (Link...but within a smaller volume of buffer that is compatible with the next solution. Add a denaturing solution...

  17. Antibody Validation Using the Indirect ELISA Method

    Type
    Protocol
    ...science. General Considerations The ELISA can be completed in a single day or broken up over several days...ELISA plate washer. Equipment Spectrophotometer compatible with 96-well plates 1–10 µL single channel pipette...

  18. Kit Free RNA Extraction

    Type
    Protocol
    ...to allow for dissociation of the nucleoprotein complexes. The effectiveness of your RNA isolation will ...

  19. Making CRISPR Plasmids Using Fragmid

    Type
    Blog Post
    ...plasmid. It shows you each component, with the ID of the component plasmid, component name, size, and any comments...module, are the components. Using the handy website, users can simply select the components they want for...able to select components that work with your design. Note that you can browse all components by clicking...Fragmid doesn’t have the components I want? You can make your own Fragmid components! Ensure your backbone... Each module of Fragmid represents a class of components, such as your CRISPR enzyme or Pol 2 promoter...’ terminus, and selection markers. Need fewer components? A spacer can be inserted instead, allowing everything...everything to stay in the correct order. Most Fragmid components are available for request on the Addgene website...

  20. 27 Hot Plasmids from 2016

    Type
    Blog Post
    ...easily design, generate, and test constructs composed of complex genetic elements. To demonstrate the usefulness...several GMAP-compatible vectors for a wide variety of biological applications. First, GMAP-compatible backbones...mammalian systems. The DULIP system is comprised of Gateway-compatible plasmids in which “bait” proteins are...plasmids designed to complement unused auxotrophies in Saccharomyces strains by compensating for histidine (... do so have been hindered by a lack of tools, compounds, and assays to monitor activation of these receptors...including a stop codon) formats. The collection is compatible with both standard Gateway cloning and the FNLCR...assembly platform for generating both simple and complex DNA constructs Have you ever wished you could ...
Showing: 141 - 160 of 950 results